U.S. patent number 6,329,381 [Application Number 09/555,292] was granted by the patent office on 2001-12-11 for heterocyclic compounds.
This patent grant is currently assigned to Japan Energy Corporation, Sumitomo Pharmaceuticals Company, Limited. Invention is credited to Hajime Kawakami, Ayumu Kurimoto, Tetsuhiro Ogino.
United States Patent |
6,329,381 |
Kurimoto , et al. |
December 11, 2001 |
Heterocyclic compounds
Abstract
The present invention relates to a heterocyclic compound of the
following general formula (I): ##STR1## wherein X is sulfur atom,
oxygen atom or --NR.sup.3 -- (R.sup.3 may form a heterocyclic ring
or a substituted heterocyclic ring with R.sup.1 via the nitrogen
atom), R.sup.1 is alkyl group, substituted alkyl group, aryl group,
substituted aryl group, heterocyclic group or substituted
heterocyclic group, and R.sup.2 is hydrogen atom, halogen atom
etc.; or its pharmaceutically acceptable salt and interferon
inducers, antiviral agents, anticancer agents and therapeutic
agents for immunologic diseases comprising the compound (I) or its
pharmaceutically acceptable salt as active ingredients.
Inventors: |
Kurimoto; Ayumu (Nishinomiya,
JP), Ogino; Tetsuhiro (Nishinomiya, JP),
Kawakami; Hajime (Nishinomiya, JP) |
Assignee: |
Sumitomo Pharmaceuticals Company,
Limited (Osaka-fu, JP)
Japan Energy Corporation (Tokyo-to, JP)
|
Family
ID: |
27341247 |
Appl.
No.: |
09/555,292 |
Filed: |
May 26, 2000 |
PCT
Filed: |
November 26, 1998 |
PCT No.: |
PCT/JP98/05318 |
371
Date: |
May 26, 2000 |
102(e)
Date: |
May 26, 2000 |
PCT
Pub. No.: |
WO99/28321 |
PCT
Pub. Date: |
June 10, 1999 |
Foreign Application Priority Data
|
|
|
|
|
Nov 28, 1997 [JP] |
|
|
9-347422 |
Dec 11, 1997 [JP] |
|
|
9-367451 |
Dec 17, 1997 [JP] |
|
|
9-367449 |
|
Current U.S.
Class: |
514/263.23;
514/263.36; 514/263.4; 544/276 |
Current CPC
Class: |
A61P
17/00 (20180101); C07D 473/24 (20130101); A61P
31/12 (20180101); A61P 37/02 (20180101); A61P
37/00 (20180101); C07D 473/18 (20130101); C07D
473/16 (20130101); A61P 35/00 (20180101); A61P
31/04 (20180101) |
Current International
Class: |
C07D
473/16 (20060101); C07D 473/18 (20060101); C07D
473/24 (20060101); C07D 473/00 (20060101); C07D
473/16 (); C07D 473/18 (); C07D 473/24 (); A61K
031/522 (); A61P 037/01 () |
Field of
Search: |
;514/262 ;544/276 |
References Cited
[Referenced By]
U.S. Patent Documents
|
|
|
6028076 |
February 2000 |
Hirota et al. |
|
Foreign Patent Documents
|
|
|
|
|
|
|
201289A2 |
|
Nov 1986 |
|
EP |
|
61-277686 |
|
Dec 1986 |
|
JP |
|
8-165292 |
|
Jun 1996 |
|
JP |
|
WO9817279 |
|
Apr 1998 |
|
JP |
|
Other References
"Field Virology, Third Ed.", B. N. Fields et al eds.,
Lippincott-Raven, Philadelphia, 1996, p. 431.* .
Testerman et al--"Journal of Leukocyte Biology", vol. 58, 365-372
(1995.) .
Stringfellow--"Methods in Enzymology", vol. 78, 262-284 (1981).
.
Good et al., The Journal of Pharmacology and Experimental
Therapeutics, vol. 227, No. 3, pp. 644-651..
|
Primary Examiner: Shah; Mukund J.
Assistant Examiner: McKenzie; Thomas C
Attorney, Agent or Firm: Birch, Stewart, Kolasch &
Birch, LLP
Parent Case Text
This application is the national phase under 35 U.S.C. .sctn.371 of
PCT International Application No. PCT/JP98/05318 which has an
International filing date of Nov. 26, 1998, which designated the
United States of America.
Claims
What is claimed is:
1. A heterocyclic compound of the following formula (I):
##STR214##
wherein X is sulfur atom, oxygen atom or --NR.sup.3a -- (in which
R.sup.3a is hydrogen atom, C.sub.1-10 alkyl group, C.sub.3-7
cycloalkyl group, or C.sub.1-10 alkyl substituted by C.sub.3-6
cyclo-alkyl group, hydroxy group, C.sub.3-6 alkoxy group, amino
group, cyano group, aryl group, aryl group substituted by C.sub.1-6
alkoxy, hydroxy or halogen, halogen atom, or nitro group, or
may form a heterocyclic ring or a substituted heterocyclic ring
together with R.sup.1 via the nitrogen atom, wherein said
substituent means C.sub.1-6 alkyl group, hydroxy C.sub.1-6 alkyl
group, C.sub.1-6 alkoxy C.sub.1-6 alkyl group, hydroxy group,
C.sub.1-6 alkoxy group, or cyano group),
R.sup.1 is C.sub.1-10 alkyl group; C.sub.3-7 cycloalkyl group;
C.sub.3-7 cycloalkyl substituted by C.sub.1-6 alkyl;
a group selected from C.sub.1-10 alkyl group, C.sub.3-7 cycloalkyl
group or C.sub.1-6 alkyl-substituted C.sub.3-7 cycloalkyl group,
which is substituted by
C.sub.3-6 cycloalkyl group, hydroxy group, C.sub.1-6 alkoxy
group,
C.sub.1-6 alkoxy group substituted by C.sub.1-6 alkoxy, hydroxy or
halogen,
amino group, C.sub.1-6 alkylamino group, cyano group, nitro group,
acyl group, carboxyl group, C.sub.2-7 alkoxycarbonyl group, halogen
atom, mercapto group, C.sub.1-6 alkylthio group,
C.sub.1-6 alkylthio group substituted by C.sub.1-6 alkoxy,
C.sub.1-6 alkylthio, hydroxy or halogen, aryl group, aryl group
substituted by C.sub.1-6 alkoxy, hydroxy or halogen, or
heterocyclic group;
aryl group;
aryl group substituted by
C.sub.1-6 alkyl group, hydroxy C.sub.1-6 alkyl group,
C.sub.1-6 alkoxy C.sub.1-6 alkyl group, hydroxy group, C.sub.1-6
alkoxy group, cyano group, amino group, C.sub.1-6 alkylamino group,
C.sub.2-7 alkoxycarbonyl group, acyl group, nitro group, halogen
atom, aryl group, aryl group substituted by C.sub.1-6 alkoxy,
hydroxy or halogen, or heterocyclic group;
heterocyclic group; or heterocyclic group substituted by
C.sub.1-6 alkyl group, hydroxy C.sub.1-6 alkyl group, C.sub.1-6
alkoxy C.sub.1-6 alkyl group, hydroxy group, C.sub.1-6 alkoxy
group, cyano group, nitro group, halogen atom, amino group,
C.sub.1-6 alkylamino group, C.sub.2-7 alkoxycarbonyl group, acyl
group, aryl group, aryl group substituted by C.sub.1-6 alkoxy,
hydroxy or halogen or
heterocyclic group, and
R.sup.2 is hydrogen atom, or one or more substituents on the
benzene ring, and said substituent is the same or different and is
hydroxy group, C.sub.1-6 alkyl group, C.sub.1-6 alkyl group
substituted by hydroxy group, C.sub.1-6 alkoxy group, carboxyl,
C.sub.2-7 alkoxycarbonyl group or halogen atom, C.sub.1-6 alkoxy
group,
C.sub.1-6 alkoxy group substituted by hydroxy group, C.sub.1-6
alkoxy group, carboxyl, C.sub.2-7 alkoxycarbonyl group or halogen
atom, C.sub.1-6 alkanoyl group,
C.sub.1-6 alkanoyl group substituted by hydroxy group, C.sub.1-6
alkoxy group, carboxyl, C.sub.2-7 alkoxycarbonyl group or halogen
atom, aroyl group, aroyl group substituted by hydroxy group,
C.sub.1-6 alkoxy group, carboxyl, C.sub.2-7 alkoxycarbonyl group or
halogen atom, carboxyl group, C.sub.2-7 alkoxycarbonyl group,
C.sub.2-7 alkoxycarbonyl group substituted by hydroxy group,
C.sub.1-6 alkoxy group, carboxyl,
C.sub.2-7 alkoxycarbonyl group or halogen atom,
amino group, C.sub.1-6 alkylamino group, di(C.sub.1-6 alkyl)amino
group, carbamoyl group, C.sub.1-6 alkylcarbamoyl group,
di(C.sub.1-6 alkyl)carbamoyl group, halogen atom, nitro group, or
cyano group,
or a pharmaceutically acceptable salt thereof.
2. The heterocyclic compound of claim 1, wherein X is sulfur atom,
or its pharmaceutically acceptable salt.
3. The heterocyclic compound of claim 1, wherein X is oxygen atom,
or its pharmaceutically acceptable salt.
4. The heterocyclic compound of claim 1, wherein X is --NH--, or
its pharmaceutically acceptable salt.
5. The heterocyclic compound of claim 1, wherein X is --NR.sup.3a
-- in which R.sup.3a means C.sub.1-6 alkyl group or substituted
C.sub.1-6 alkyl substituted by
C.sub.3-6 cycloalkyl group, hydroxy group, C.sub.1-6 alkoxy
group,
amino group, cyano group, aryl group,
aryl group substituted by C.sub.1-6 alkoxy, hydroxy or halogen,
halogen atom, or nitro group,
or its pharmaceutically acceptable salt.
6. The heterocyclic compound of claim 1, wherein R.sup.3a forms a
heterocyclic ring or a substituted heterocyclic ring together with
R.sup.1 via the nitrogen atom, or its pharmaceutically acceptable
salt.
7. The heterocyclic compound of claim 1, wherein R.sup.1 means
C.sub.1-6 alkyl group or substituted C.sub.1-6 alkoxy, C.sub.1-6
alkylthio, C.sub.2-7 alkoxycarbonyl, hydroxy, halogen atom, cyano,
amino, cyclohexyl, trifluoromethyl, pyridyl, phenyl, methoxyphenyl,
hydroxyphenyl, halophenyl or thienyl, or its pharmaceutically
acceptable salt.
8. The heterocyclic compound of claim 1, wherein R.sup.1 means
C.sub.1-6 substituted alkyl group and said substituent is hydroxyl,
or its pharmaceutically acceptable salt.
9. 6-Amino-9-benzyl-8-hydroxy-2-[(2-hydroxyethyl)thio]purine or its
pharmaceutically acceptable salt.
10. 6-Amino-9-(4-fluorobenzyl)-8-hydroxy-2-(2-methoxyethoxy)purine
or its pharmaceutically acceptable salt.
11. 6-Amino-9-benzyl-8-hydroxy-2-(2-methoxyethoxy)purine or its
pharmaceutically acceptable salt.
12. A pharmaceutical composition comprising a heterocyclic compound
or its pharmaceutically acceptable salt of claim 1, as an active
ingredient.
13. A method for inducing interferons in a patient having virus
infected disease, cancer or allergic disease by administering the
heterocyclic compound of claim 1, or its pharmaceutically
acceptable salt to the patient in an amount effective to induce
interferons.
Description
TECHNICAL FIELD
The present invention relates to novel heterocyclic compounds
having inducing activity for biosynthesis of interferon. The
heterocyclic compounds of the present invention induce biosynthesis
of endogenous interferon in a living body, and are useful for
medicines, such as antiviral agents, anticancer agents and
therapeutic agents for immunologic diseases.
BACKGROUND OF THE ART
It has been recently determined that endogenous interferon plays
not only central role to bio-defensive mechanism against virus
infections and microbial infections, but also an important role dn
antitumor and immune modulator. Mass production of interferon is
established. Namely, it is possible to obtain of natural interferon
by cell culture and also to produce a large amount of recombinant
interferon from E. coli transferred with a gene of interferon and
therefore, many research achievements on these interferons have
accumulated. For example, many kinds of biological activity on
interferon, such as antiviral activity, prevention of cell growth
and immune modulation have been confirmed and interferon is
practiced on clinics as treating agents for virus infected
diseases, such as hepatitis C and hepatitis B, anticancer agents
and therapeutic agents for immunologic disease. Furthermore, it is
suggested that interferon will prevent carcinogenesis by hepatitis
C and hepatitis B.
Since there is no therapeutic method for almost of the above
diseases, interferon is especially made much of.
DISCLOSURE OF INVENTION
The object of the present invention is to provide novel low
molecular weight compounds having inducing activity for
biosynthesis of interferon, and interferon inducers, antiviral
agents, anticancer agents and therapeutic agents for immunologic
diseases comprising these compounds as active ingredients.
Viruses of many kinds of animals, microbes such as mycobacteria and
protozoa, extracts of them, mitogen, specific antigens and
immunopotenciators are known as inducers for biosynthesis of
interferon. It is known that for example, many kinds of natural
double strand RNAS, synthesized double strand RNAs such as
poly-I:C, and anionic high molecular compounds such as polyacrylic
acid and oxyamylose oxidized with chlorite have inductive activity
of interferon.
On the other hand, among low molecular weight compounds have been
found fluorenones, pyrimidine derivatives, anthraquinones,
acridines and so on having inductive activity of interferon
(Stringfollow, D. A.: Methods in Enzymology, 1981, 78, 262).
However, when these compounds are used in clinical trial, their
inducing activity of interferon is unexpectedly low and these
compounds have side effects or by administering them repeatedly,
their inducing activity of interferon decreases and therefore,
development on these compounds has not succeeded. Furthermore,
imidazo-quinolines are known as interferon inducers among low
molecular compounds. However, it is known that these compounds are
inferior in selective interferon inducing activity and
simultaneously induce cytokines such as IL-6, TNF-.alpha., etc
(Testerman, T. L., et al.: J. Leukocyte Biol., 1995, 58, 365).
As the result of extensive investigation of interferon biosynthesis
inducers among low molecules, the present inventors have found that
the heterocyclic compounds of the present invention have excellent
interferon biosynthesis inducing activity.
The present invention relates to a heterocyclic compound of the
following general formula (I): ##STR2##
wherein X is sulfur atom, oxygen atom, --NR.sup.3 -- (in which
R.sup.3 is hydrogen atom, alkyl group or substituted alkyl, or may
form a heterocyclic ring or a substituted heterocyclic ring
together with R.sup.1 via the nitrogen atom),
R.sup.1 is alkyl group, substituted alkyl group, aryl group,
substituted aryl group, heterocyclic group or substituted
heterocyclic group, and
R.sup.2 is hydrogen atom, or one or more substituents on the
benzene ring, and said substituent is the same or different and is
hydroxy group, lower alkyl group, substituted lower alkyl group,
lower alkoxy group, substituted lower alkoxy group, lower alkanoyl
group, substituted lower alkanoyl group, aroyl group, substituted
aroyl group, carboxyl group, lower alkoxycarbonyl group,
substituted lower alkoxycarbonyl group, amino group, lower
alkylamino group, di(lower alkyl)amino group, carbamoyl group,
lower alkylcarbamoyl group, di(lower alkyl)carbamoyl group, halogen
atom, nitro group or cyano group; or a pharmaceutically acceptable
salt thereof.
Further, the present invention relates to a pharmaceutical
composition comprising a heterocyclic compound of the above formula
(I) or its pharmaceutically acceptable salt as an active
ingredient.
Further, the present invention relates to an interferon inducer, an
antiviral agent, an anticancer agent and a therapeutic agent for
immunologic diseases comprising a heterocyclic compound of the
above formula (I) or its pharmaceutically acceptable salt as an
active ingredient.
Furthermore, the present invention relates to a process for
preparing a heterocyclic compound of the above formula (I) or its
pharmaceutically acceptable salt.
Groups R.sup.1, R.sup.2 and R.sup.3 in the formula (I) are
explained below.
In R.sup.1 alkyl group includes straight or branched C.sub.1-10
alkyl group (e.g. methyl, ethyl, propyl, butyl, pentyl, isopropyl,
isobutyl, 1-methylpropyl, 3-methylbutyl or hexyl), C.sub.3-7
cycloalkyl group (e.g. cyclopropyl, cyclopentyl, cyclohexyl or
cycloheptyl), and alkyl-substituted C.sub.3-7 cycloalkyl group,
preferably straight or branched C.sub.1-6 alkyl group (e.g. methyl,
ethyl, propyl, butyl or pentyl), and C.sub.5-7 cycloalkyl group
(e.g. cyclopentyl or cyclohexyl).
In R.sup.1 substituted alkyl group means the above alkyl
substituted by the same or different and one or more
substituents.
Said substituents include cycloalkyl group (C.sub.3-6 cycloalkyl
group, such as cyclopropyl, cyclopentyl or cyclohexyl), hydroxy
group, lower alkoxy group (C.sub.1-6 alkoxy such as methoxy,
ethoxy, propoxy, butoxy or pentoxy), substituted lower alkoxy group
(substituted C.sub.1-6 alkoxy group, such as methoxyethoxy,
ethoxyethoxy, hydroxyethoxy or chloroethoxy), amino group,
alkylamino group, cyano group, nitro group, acyl group, carboxyl
group, lower alkoxycarbonyl group (C.sub.2-7 alkoxycarbonyl group,
such as methoxycarbonyl or ethoxycarbonyl), halogen atom, such as
fluorine atom, chlorine atom or bromine atom, mercapt group, lower
alkylthio group (C.sub.1-6 alkylthio group, such as methylthio,
ethylthio, propylthio or butylthio), substituted lower alkylthio
group (substituted C.sub.1-6 alkylthio group, such as
methoxyethylthio, methylthioethylthio, hydroxyethylthio or
chloroethylthio), aryl group (C.sub.6-10 monocyclic or fused cyclic
aryl group, such as phenyl or naphthyl), substituted aryl group
(substituted C.sub.6-10 monocyclic or fused cyclic aryl group, such
as 4-hydroxyphenyl, 4-methoxyphenyl, 4-fluorophenyl, 4-chlorophenyl
or 3,4-dichlorophenyl), and heterocyclic group (5-6 membered
saturated heterocyclic group containing nitrogen atoms from 0-2 and
oxygen atoms from 0-2, such as piperidyl, piperazinyl, morpholinyl,
tetrahydrofuranyl, pyrrolidinyl, pyrazolinyl or 1,3-dioxolanyl, 5-6
membered unsaturated heterocyclic group, such as furyl, pyrrolyl,
pyrazolyl, imidazolyl, thiazolyl, thienyl, pyridyl or pyrimidinyl,
or bicyclic unsaturated heterocyclic group, such as indolyl,
isoindolyl, quinolyl, benzothiazolyl, chromanyl, benzofuranyl or
phthalimino).
In R.sup.1 aryl group means C.sub.6-10 monocyclic or fused cyclic
aryl group, such as phenyl or naphthyl.
In R.sup.1 substituted aryl group means the above aryl group
substituted by the same or different and one or more
substituents.
Said substituent includes lower alkyl group (C.sub.1-6 alkyl group,
such as methyl, ethyl, propyl, butyl, cyclopentyl or cyclohexyl),
hydroxy lower alkyl group (hydroxy C.sub.1-6 alkyl group, such as
hydroxymethyl, 2-hydroxyethyl or 3-hydroxypropyl), lower alkoxy
lower alkyl group (C.sub.1-6 alkoxy C.sub.1-6 alkyl group, such as
2-methoxyethyl, 2-ethoxyethyl or 3-methoxypropyl), hydroxy group,
lower alkoxy group (C.sub.1-6 alkoxy group, such as methoxy,
ethoxy, propoxy, butoxy or pentoxy), cyano group, amino group,
substituted amino group, lower alkoxycarbonyl group (C.sub.2-7
alkoxycarbonyl group, such as methoxycarbonyl, ethoxycarbonyl or
propoxycarbonyl), acyl group, nitro group, halogen atom, such as
fluorine atom, chlorine atom or bromine atom, aryl group
(C.sub.6-10 monocyclic or fused cyclic aryl group, such as phenyl
or naphthyl), substituted aryl group (substituted C.sub.6-10
monocyclic or fused cyclic aryl group, such as 4-hydroxyphenyl,
4-methoxyphenyl, 4-chlorophenyl or 3,4-dichlorophenyl), and
heterocyclic group (alicyclic or aromatic heterocyclic group
containing nitrogen atoms from 1-2 and oxygen atom from 0-1, such
as pyrrolidinyl, piperidyl, piperazinyl or morpholinyl).
In R.sup.1 heterocyclic group means monocyclic saturated
heterocyclic group, or unsaturated monocyclic or fused heterocyclic
group containing at least one heteroatom, that is, 0-3 nitrogen
atoms, 0-1 oxygen atom and 0-1 sulfur atom.
Said saturated monocyclic heterocyclic group includes 5 or 6
membered saturated heterocyclic group, such as tetrahydrofuranyl,
pyrrolidinyl, morpholinyl, piperidyl, piperazinyl or pyrazolidinyl.
Said unsaturated monocyclic heterocyclic group means 5 or 6
membered unsaturated heterocyclic group, such as furyl, pyrrolyl,
pyrazolyl, imidazolyl, thiazolyl, thienyl, pyridyl or pyrimidinyl.
Said unsaturated fused heterocyclic group means unsaturated
bicyclic heterocyclic group, such as indolyl, isoindolyl, quinolyl,
benzothizolyl, chromanyl or benzofuranyl.
In R.sup.1 substituted heterocyclic group means the above
heterocyclic group substituted by the same or different and one or
more substituents.
Said substituents include lower alkyl group (C.sub.1-6 alkyl group,
such as methyl, ethyl, propyl, butyl, cyclopentyl or cyclohexyl),
hydroxy lower alkyl group (hydroxy C.sub.1-6 alkyl group, such as
hydroxymethyl, 2-hydroxyethyl or 3-hydroxypropyl), lower alkoxy
lower alkyl group (C.sub.1-6 alkoxy C.sub.1-6 alkyl group, such as
2-methoxyethyl, 2-ethoxyethyl or 3-methoxypropyl), hydroxy group,
lower alkoxy group (C.sub.1-6 alkoxy group such as methoxy, ethoxy,
propoxy, butoxy or pentoxy), cyano group, nitro group, halogen
atom, such as fluorine atom, chlorine atom or bromine atom, amino
group, substituted amino group, lower alkoxycarbonyl group
(C.sub.2-7 alkoxycarbonyl group, such as methoxycarbonyl,
ethoxycarbonyl or propoxycarbonyl), acyl group, aryl group
(C.sub.6-10 monocyclic or fused cyclic aryl group, such as phenyl
or naphthyl), substituted aryl group (substituted C.sub.6-10
monocyclic or fused cyclic aryl group, such as 4-hydroxyphenyl,
4-methoxyphenyl, 4-chlorophenyl or 3,4-dichlorophenyl), and
heterocyclic group (alicyclic or aromatic heterocyclic group
containing nitrogen atoms from 1-2 and oxygen atom from 0-1, such
as pyrrolidinyl, piperidyl, piperazinyl or morpholinyl).
In R.sup.2 lower alkyl group includes C.sub.1-6 alkyl group (e.g.
methyl, ethyl, propyl, 1-methylethyl, butyl, 1-methylpropyl,
2-methylpropyl, 1,1-dimethylethyl, pentyl, 1-methylbutyl,
2-methylbutyl, 3-methylbutyl, 1,1-dimethylpropyl,
1,2-dimethylpropyl, 2,2-dimethylpropyl).
In R.sup.2 substituted lower alkyl group means the above alkyl
substituted by the same or different and one or more
substituents.
Said substituents include hydroxy group, lower alkoxy group (for
example, C.sub.1-6 alkoxy group, such as methoxy, ethoxy or
propoxy), carboxyl group, lower alkoxycarbonyl group (for example,
C.sub.2-7 alkoxycarbonyl group, such as methoxycarbonyl,
ethoxycarbonyl or propoxycarbonyl) and halogen atom, such as
fluorine atom, chlorine atom or bromine atom.
In R.sup.2 lower alkoxy group means C.sub.1-6 alkoxy group, such as
methoxy, ethoxy or propoxy.
In R.sup.2 substituted lower alkoxy group means the above alkoxy
group substituted by the same or different and one or more
substituents.
Said substituents include hydroxy group, lower alkoxy group
(C.sub.1-6 alkoxy group, such as methoxy, ethoxy or propoxy),
carboxyl group, lower alkoxycarbonyl group (C.sub.2-7
alkoxy-carbonyl group, such as methoxycarbonyl, ethoxycarbonyl
group or propoxycarbonyl) and halogen atom, such as fluorine atom,
chlorine atom or bromine atom.
In R.sup.2 lower alkanoyl group means C.sub.1-6 alkanoyl group,
such as formyl, acetyl, propanoyl, butanoyl, pentanoyl or
hexanoyl.
In R.sup.2 substituted lower alkanoyl group means the above
alkanoyl group substituted by the same or different and one or more
substituents.
Said substituents include hydroxy group, lower alkoxy group
(C.sub.1-6 alkoxy group, such as methoxy, ethoxy or propoxy),
carboxyl group, lower alkoxycarbonyl group (C.sub.2-7
alkoxycarbonyl group, such as methoxycarbonyl or propoxycarbonyl)
and halogen atom, such as fluorine atom, chlorine atom or bromine
atom.
In R.sup.2 aroyl group means C.sub.7-11 aroyl group, such as
benzoyl or naphthoyl.
In R.sup.2 substituted aroyl group means the above aroyl group
substituted by the same or different and one or more
substituents.
Said substituents include hydroxy group, lower alkoxy group
(C.sub.1-6 alkoxy group, such as methoxy, ethoxy or propoxy),
carboxyl group, lower alkoxycarbonyl group (C.sub.2-7
alkoxycarbonyl group, such as methoxycarbonyl, ethoxycarbonyl or
propoxycarbonyl) and halogen atom, such as fluorine atom, chlorine
atom or bromine atom.
In R.sup.2 lower alkoxycarbonyl group means C.sub.2-7
alkoxycarbonyl group, such as methoxycarbonyl, ethoxycarbonyl or
propoxycarbonyl.
In R.sup.2 substituted lower alkoxycarbonyl group means the above
alkoxycarbonyl group substituted by the same or different and one
or more substituents.
Said substituents include hydroxy group, lower alkoxy group
(C.sub.1-6 alkoxy group, such as methoxy, ethoxy or propoxy),
carboxyl group, lower alkoxycarbonyl group (C.sub.2-7
alkoxycarbonyl group, such as methoxycarbonyl, ethoxycarbonyl or
propoxycarbonyl) and halogen atom, such as fluorine atom, chlorine
atom or bromine atom.
In R.sup.2 lower alkylamino group means amino group substituted by
C.sub.1-6 alkyl group (e.g. methylamino, ethylamino, propylamino,
butylamino).
In R.sup.2 di(lower alkyl)amino group means amino group substituted
by the same or different and C.sub.1-6 alkyl group (e.g.
dimethylamino, diethylamino, ethylmethylamino).
In R.sup.2 lower alkylcarbamoyl group means carbamoyl group
substituted by C.sub.1-6 alkyl group (e.g. methylcarbamoyl,
ethylcarbamoyl, propylcarbamoyl, butylcarbamoyl).
In R.sup.2 di(lower alkyl)carbamoyl group means carbamoyl group
substituted by the same or different and C.sub.1-6 alkyl group
(e.g. dimethylcarbamoyl, diethylcarbamoyl,
ethylmethylcarbamoyl).
In R.sup.2 halogen atom means halogen atom such as fluorine atom,
chlorine atom, bromine atom or iodine atom.
In R.sup.3 alkyl group includes straight or branched C.sub.1-10
alkyl group (e.g. methyl, ethyl, propyl, butyl, pentyl, hexyl) and
C.sub.3-7 cycloalkyl group (e.g. cyclopropyl, cyclopentyl,
cyclohexyl, cycloheptyl), preferably straight or branched C.sub.1-6
alkyl group (e.g. methyl, ethyl, propyl, butyl, pentyl), and
C.sub.5-7 cycloalkyl group (e.g. cyclopentyl, cyclohexyl).
In R.sup.3 substituted alkyl group means the above alkyl group
substituted by the same or different and one or more
substituents.
Said substituents include cycloalkyl group (C.sub.3-6 cycloalkyl
group, such as cyclopropyl, cyclopentyl or cyclohexyl), hydroxy
group, lower alkoxy group (C.sub.1-6 alkoxy, such as methoxy,
ethoxy, propoxy, butoxy or pentoxy), amino group, cyano group, aryl
group such as phenyl, substituted aryl group, such as
4-hydroxyphenyl, 4-methoxyphenyl, 4-chlorophenyl or
3,4-dichlorophenyl, nitro group and halogen atom, such as fluorine
atom, chlorine atom or bromine atom.
Heterocyclic ring formed together with R.sup.3 and R.sup.1 via the
nitrogen atom means 5 or 6 membered saturated heterocyclic ring,
such as 1-pyrrolidinyl, 4-morpholinyl, 1-piperidyl, 1-piperazinyl
or 1-pyrazolidinyl, and 5 or 6 membered unsaturated heterocyclic
ring such as 1-imidazolyl.
Substituted heterocyclic ring formed together with R.sup.3 and
R.sup.1 via the nitrogen atom means the above heterocyclic ring
formed together with R.sup.3 and R.sup.1 via the nitrogen atom
alkyl substituted by the same or different and one or more
substituents.
Said substituents include lower alkyl group (C.sub.1-6 alkyl group,
such as methyl, ethyl, propyl, butyl, cyclopentyl or cyclohexyl),
hydroxy lower alkyl group (hydroxy C.sub.1-6 alkyl, such as
hydroxymethyl, 2-hydroxyethyl or 3-hydroxypropyl), lower alkoxy
lower alkyl group (C.sub.1-6 alkoxy C.sub.1-6 alkyl, such as
2-methoxyethyl, 2-ethoxyethyl or 3-methoxypropyl), hydroxy group,
lower alkoxy group (C.sub.1-6 alkoxy, such as methoxy, ethoxy,
propoxy, butoxy or pentoxy) and cyano group.
The compound (I) of the present invention forms an equilibrium
mixture with a tautomer represented by the following formula (Ia):
##STR3##
The compound (I) of the present invention may forms a salt with an
acid.
The preferable acids are pharmaceutically acceptable acids,
including inorganic acids, such as hydrochloric acid, sulfuric
acid, hydrobromic acid, etc., organic acids, such as acetic acid,
oxalic acid, citric acid, malic acid, tartaric acid, fumaric acid,
maleic acid, etc.
Further, in case of the compound (I) having an acidic substituent,
the compound may form a salt with a base.
The preferable bases are pharmaceutically acceptable bases,
including inorganic bases like alkali metals, such as sodium or
potassium, or organic bases, such as triethylamine or pyridine.
Preferable embodiments among the compounds (I) of the present
invention are as follows.
(a) A heterocyclic compound of the formula (II): ##STR4##
wherein X.sup.1 is sulfur atom, oxygen atom or --NR.sup.3a -- in
which R.sup.3a is hydrogen, C.sub.1-6 alkyl group, or substituted
C.sub.1-6 alkyl group, or may form a heterocyclic ring or a
substituted heterocyclic ring together with R.sup.1a via the
nitrogen atom,
R.sup.1a is C.sub.1-6 alkyl group, substituted C.sub.1-6 alkyl
group, aryl group, substituted aryl group, heterocyclic group or
substituted heterocyclic group, and
R.sup.2a is hydrogen atom, or one or more substituents on the
benzene ring, and said substituent is the same or different and is
halogen atom, C.sub.1-6 alkoxy group, nitro group or hydroxy group;
or
its pharmaceutically acceptable salt.
(b) A heterocyclic compound of the above (a) wherein X.sup.1 is
sulfur atom.
(c) A heterocyclic compound of the above (a) wherein X.sup.1 is
oxygen atom.
(d) A heterocyclic compound of the above (a) wherein X.sup.1 is
--NH--.
(e) A heterocyclic compound of the above (a) wherein X.sup.1 is
--NR.sup.3a -- in which R.sup.3a means C.sub.1-6 alkyl group or
substituted C.sub.1-6 alkyl group.
(f) A heterocyclic compound of the above (a) wherein R.sup.3a forms
a heterocyclic ring or a substituted heterocyclic ring together
with R.sup.1a via the nitrogen atom.
(g) A heterocyclic compound of any of the above (a)-(d) wherein
R.sup.1a means C.sub.1-6 alkyl group or substituted C.sub.1-6 alkyl
group.
(h) A heterocyclic compound of any of the above (a)-(d) wherein
R.sup.1a means C.sub.1-6 alkyl group substituted by C.sub.1-6
alkoxy, hydroxy, halogen atom, cyano, trifluoromethyl, pyridyl,
phenyl, tolyl or thienyl.
(i) A heterocyclic compound of any of the above (a)-(d) wherein
R.sup.1a means C.sub.1-6 alkyl group.
(j) A heterocyclic compound of any of the above (a)-(d) wherein
R.sup.1a means C.sub.3-6 cycloalkyl group.
(k) A pharmaceutically acceptable salt of a heterocyclic compound
of any of the above (b)-(j).
The compound (I) of the present invention has excellent inducing
activity for biosynthesis of interferon and shows in general the
following structure activity-relationship between R.sup.1 and
R.sup.2 of the compound (I).
The inducing activity for biosynthesis of the compound (I) of the
present invention is influenced by length or size of R.sup.1. For
instance, the activity at the minimum concentration reaches best
when carbon number on the group of R.sup.1 is around 3 or 4 and
therefore, it shows bell-type activity-relationship.
On the other hand, although the activity at the minimum
concentration when carbon number on the group of R.sup.1 is around
1 or 2 is inferior to the activity when carbon number on the group
of R.sup.1 is around 3 or 4, the former is superior to the latter
in induction amount of interferon judging from the view of the
production of interferon.
Therefore, when R.sup.1 is alkyl group, preferable range of carbon
number in the view of the activity is 1 to 8, more preferably 3 to
5.
Preferable embodiments of alkyl group in R.sup.1 are methyl, ethyl,
propyl, butyl, pentyl, isopropyl, isobutyl, 1-methylpropyl,
3-methylbutyl, cyclopentyl and cyclohexyl.
Furthermore, it has been made clear that in case that R.sup.1 is
substituted alkyl group, the activity is also influenced by size or
length of R.sup.1.
That is, the activity is influenced by total size or length of
R.sup.1 including a substituent (e.g. lower alkoxy, hydroxy,
halogen atom). For instance, preferable range of length in the view
of the activity is 1 to 8, more preferably 3 to 5 by calculating in
carbon number as the same as in alkyl group in R.sup.1.
Preferable embodiment of alkyl group in R.sup.1 are 2-hydroxyethyl,
3-hydroxypropyl, 4-hydroxybutyl, 2-aminoethyl, 3-aminopropyl,
4-aminobutyl, methoxymethyl, 2-methoxyethyl, 3-methoxypropyl,
ethoxymethyl, 2-ethoxyethyl, methylthiomethyl, 2-methylthioethyl,
3-methylthiopropyl, 2-fluoroethyl, 3-fluoropropyl,
2,2,2-trifluoroethyl, cyanomethyl, 2-cyanoethyl, 3-cyanopropyl,
methoxycarbonylmethyl, 2-methoxycarbonylethyl,
3-methoxycarbonylpropyl, benzyl, phenethyl, 4-pyridylmethyl,
cyclohexylmethyl, 2-thienylmethyl, 4-methoxyphenylmethyl,
4-hydroxyphenylmethyl, 4-fluorophenylmethyl, and
4-chlorophenylmethyl.
When R.sup.1 is aryl group or substituted aryl group, the same
tendency as above is observed. The most preferable embodiment of
aryl group or substituted aryl group in R.sup.1 are phenyl,
4-methoxyphenyl, 4-hydroxyphenyl, 4-fluorophenyl and
4-chlorophenyl.
When R.sup.1 is heterocyclic group or substituted heterocyclic
group, or R.sup.3 forms heterocyclic ring or substituted
heterocyclic ring with R.sup.1, the same tendency as above is also
observed. The most preferable embodiments of heterocyclic group or
substituted heterocyclic group in R.sup.1 are 1-pyrrolidinyl,
4-morpholinyl and 3-(2-hydroxyethyl)-1-pyrrolidinyl.
Preferable X are sulfur atom, oxygen atom and nitrogen atom which
may be substituted, in order.
Although influence by R.sup.2 on inducing activity for biosynthesis
of interferon is not so remarkable as that by R.sup.1, preferable
embodiments of R.sup.2 are hydrogen atom, halogen atom such as
fluorine atom or chlorine atom, hydroxy group, lower alkoxy group
such as methoxy, and nitro group.
A preferable substituent of R.sup.2 and its position among the
above embodiments is 4-fluoro, 4-chloro, 4-hydroxy, 4-methoxy,
4-nitro, 2,4-difluoro, 2,4-dichloro, 3,4-difluoro, 3,4-dichloro, or
3,4-dimethoxy.
Process for preparation of the compound of the present
invention.
The compound of the present invention can be prepared by the
following methods. However, starting materials which are not
described below are prepared in accordance with the following
methods, known methods, or in accordance with known methods.
PROCESS 1
##STR5##
wherein R.sup.1, X and R.sup.2 are the same as definition in the
formula (I), Y is leaving group such as halogen atom (e.g. chlorine
atom, bromine atom), R.sup.5 is alkyl group, and R.sup.a and
R.sup.b are hydrogen atom, or mean amino protective group because
they are protected by an amino protective group on the way of
reaction, if necessary.
Compound (3) is prepared by reacting compound (2) with NHR.sup.a
R.sup.b in an aqueous solution or in an organic solvent.
NHR.sup.a R.sup.b can be used about equal molar or large excess
amount to compound (2).
Organic solvents are alcohols, such as methanol, ethanol, propanol
or butanol, ethers such as tetrahydrofuran, 1,4-dioxane or diglyme,
or aprotic solvents, such as dimethylformamide, dimethyl sulfoxide,
acetonitrile or hexamethylphosphoroustriamide [(CH.sub.3).sub.2
N).sub.3 P].
The reaction temperature is selected from the range between room
temperature and about 200.degree. C.
Reaction vessels such as an autoclave etc. may be used in the
reaction, if necessary.
Compound (5) is prepared by reacting compound (3) and compound (4)
in the presence of a base in an organic solvent.
Compound (4) can be used about equal molar or several molars to
compound (3).
Bases are inorganic bases such as alkali metal carbonates (e.g.
sodium carbonate, potassium carbonate), or organic bases, such as
tertiary amines (e.g. triethylamine, diisopropylethylamine) or
pyridines (e.g. 4-dimethylaminopyridine, pyridine). The base is
preferably used about equimolar to compound (4)
The organic solvents are halogenated hydrocarbons such as
tetrachloromethane, chloroform or methylene chloride, ethers such
as diethyl ether, tetrahydrofuran or 1,4-dioxane, or aprotic
solvents, such as dimethylformamide, dimethyl sulfoxide,
acetonitrile or hexamethylphosphoroustriamide.
The reaction temperature is selected from the range between about
0.degree. C. and around the boiling point of the solvent.
Compound (6) is prepared by reacting compound (5) with Br.sub.2 in
an organic solvent.
A reaction promoter such as sodium acetate may be added to the
reaction mixture.
Br.sub.2 is used from equimolar to several moles of compound (5),
preferably from equimolar to one and one-half moles.
The organic solvents are halogenated hydrocarbons, such as
tetrachloromethane, chloroform or methylene chloride, ethers, such
as diethyl ether, acetic acid, or carbon disulfide.
The reaction temperature is selected from the range between about
0.degree. C. and around boiling point of the solvent.
Compound (7) is prepared by reacting compound (6) and an alcohol
such as methanol in the presence of a base in an organic
solvent.
The base are alkali metals, such as sodium or potassium, alkali
metal hydrides, such as sodium hydride or potassium hydride,
organometallic compounds, such as methyl lithium, butyl lithium or
lithium diisopropylamide.
The base is preferably used from about equal molar to about two
times as much to compound (6).
The organic solvents are ethers, such as diethyl ether,
tetrahydrofuran or 1,4-dioxane, or aprotic solvents, such as
dimethylformamide, dimethyl sulfoxide, acetonitrile or
hexamethylphosphoroustriamide. The alcohol as the reagent, such as
methanol, ethanol, propanol or butanol may serve as a solvent.
The reaction temperature is selected from the range between about
room temperature and around boiling point of the solvent.
Compound (8) is prepared by reacting compound (7) with R.sup.1 XH
in an organic solvent.
R.sup.1 XH is used from about equal molar to several molars to
compound (7).
When X is oxygen atom or sulfur atom, the reaction is preferably
carried out in the presence of a base.
The bases are alkali metals, such as sodium or potassium, alkali
metal hydrides, such as sodium hydride or potassium hydride,
organometalic compounds, such as methyl lithium, butyl lithium or
lithium diisopropylamide. The base is preferably used about
equimolar to R.sup.1 XH.
The organic solvents are aprotic solvents, such as
dimethylformamide, acetonitrile or hexamethylphosphoroustriamide,
or ethers such as diethyl ether, tetrahydrofuran or 1,4-dioxane or
diglyme.
The reaction temperature is selected from the range between about
room temperature and around the boiling point of the solvent.
Compound (9) is prepared by treating compound (8) with an acid in
water or a mixture of water and an organic solvent.
The acids are inorganic acids, such as hydrochloric acid or
hydrobromic acid, or organic acids such as trifluoroacetic
acid.
The organic solvents are ethers, such as diethyl ether or
tetrahydrofuran, aprotic solvents such as dimethylformamide,
alcohols, such as methanol, ethanol or propanol, or acetic
acid.
The reaction temperature is selected from the range between about
room temperature and around boiling point of the solvent.
PROCESS 2
##STR6##
wherein R.sup.1, X and R.sup.2 are the same as definition in the
formula (I), Z is halogen atom such as chlorine atom or bromine
atom, or leaving group such as methanesulfonyloxy or
p-toluenesulfonyloxy, and Y and R.sup.5 are the same as defined
above.
Compound (11) is prepared by a method known by the skilled person.
For instance, when Z is a chlorine atom, compound (11) is prepared
by reacting compound (10) with phosphorousoxychloride.
The reaction temperature is selected from the range between room
temperature and reflux temperature of the reaction solvent. When Z
is methanesulfonyloxy, compound (11) is prepared by reacting
compound (10) with methanesulfonyl chloride in the presence of a
base in an organic solvent, and if necessary, NH.sub.2 group on
compound (10) is protected and then deprotected.
The bases are inorganic bases such as alkali metal carbonates (e.g.
potassium carbonate), or organic bases, such as triethylamine,
diisopropylethylamine, 4-dimethylaminopyridine or pyridine.
The organic solvents are halogenated hydrocarbons such as methylene
chloride, ethers, such as diethyl ether or tetrahydrofuran, or
aprotic solvents such as dimethylformamide, etc.
The reaction temperature is selected from the range between about
0.degree. C. and around boiling point of the solvent.
Compound (12) is prepared by reacting compound (11) with R.sup.1 XH
in an organic solvent.
When X is oxygen atom or sulfur atom, the reaction is preferably
carried out in the presence of a base. The bases are alkali metals,
such as sodium or potassium, alkali metal hydrides, such as sodium
hydride or potassium hydride, or organometalic compounds, such as
methyl lithium, butyl lithium or lithium diisopropylamide.
The organic solvents are aprotic solvents, such as
dimethylformamide, acetonitrile or hexamethylphosphoroustriamide,
or ethers, such as diethyl ether, tetrahydrofuran 1,4-dioxane or
diglyme.
The reaction temperature is selected from the range between about
room temperature and around the boiling point of the solvent.
Compound (13) is prepared by reacting compound (12) and compound
(4) in the presence of a base in an organic solvent.
The bases are inorganic bases such as alkali metal carbonates (e.g.
sodium carbonate, potassium carbonate), or organic bases, such as
tertiary amines (e.g. triethylamine, diisopropylethylamine) or
pyridines (e.g. dimethylaminopyridine, pyridine).
The organic solvents are halogenated hydrocarbons such as methylene
chloride etc., ethers, such as diethyl ether or tetrahydrofuran, or
aprotic solvents, such as dimethylformamide, dimethyl sulfoxide or
acetonitrile.
The reaction temperature is selected from the range between about
room temperature and around the boiling point of the solvent.
Compound (14) is prepared by nitrating compound (13) in an organic
solvent, for example by adding nitric acid thereto in an organic
solvent such as acetic acid.
The reaction temperature is selected from the range between about
-20.degree. C. and around the boiling point of the solvent.
Compound (15) is prepared by reducing nitro group on compound (14)
in an organic solvent.
The reducing agents are hydrogen, sodium borohydride or lithium
aluminum hydride.
The organic solvents are alcohols, such as methanol or ethanol,
esters such as ethyl acetate, etc., or ethers, such as diethyl
ether or tetrahydrofuran.
The reaction temperature is selected from the range between about
0.degree. C. and around the boiling point of the solvent.
Compound (16) is prepared by reacting compound (15) with formic
acid or trimethyl orthoformate in the presence of an acid.
The acids are inorganic acids such as hydrochloric acid, or organic
acids, such as p-toluenesulfonic acid or camphor sulfonic acid.
The reaction temperature is selected from the range between about
room temperature and around the boiling point of the solvent.
Compound (17) is prepared by reacting compound (16) and Br.sub.2 in
an organic solvent.
A reaction promoter such as sodium acetate may be added in this
reaction.
The organic solvents are halogenated hydrocarbons, such as
tetrachloromethane, methylene chloride or dichloroethane, ethers
such as diethyl ether, acetic acid, or carbon disulfide.
The reaction temperature is selected from the range between about
0.degree. C. and around the boiling point of the solvent.
Compound (18) is prepared by reacting compound (17) and R.sup.5 OH
in the presence of a base in an organic solvent.
The bases are alkali metals, such as sodium or potassium, alkali
metal hydrides, such as sodium hydride or potassium hydride, or
organometalic compounds, such as methyl lithium, butyl lithium or
lithium diisopropylamide.
The organic solvents are ethers, such as diethyl ether or
tetrahydrofuran, or aprotic solvents, such as dimethylformamide or
acetonitrile. The alcohol used as the reagent, such as methanol,
ethanol, propanol or butanol may be served as a solvent.
The reaction temperature is selected from the range between about
room temperature and around the boiling point of the solvent.
Compound (19) is prepared by treating compound (18) with an acid in
water or a mixture of water and an organic solvent.
The acids are inorganic acids, such as hydrochloric acid or
hydrobromic acid, or organic acids such as trifluoroacetic acid,
etc.
The organic solvents are ethers, such as diethyl ether or
tetrahydrofuran, aprotic solvents, such as dimethylformamide or
acetonitrile, alcohols, such as methanol, ethanol or propanol, or
acetic acid.
The reaction temperature is selected from the range between about
room temperature and around the boiling point of the solvent.
PROCESS 3
##STR7##
wherein R.sup.1, X and R.sup.2 are the same as definition in the
formula (I), and Y is the same as defined above.
Compound (22) is prepared by reacting compound (20) and compound
(21) in an organic solvent. The reaction can be carried out in the
presence or absence of a solvent.
The bases are inorganic bases such as alkali metal carbonates (e.g.
potassium carbonate), or organic bases, such as tertiary amines
(e.g. triethylamine, diisopropylethylamine) or pyridines (e.g.
4-dimethylaminopyridine, pyridine).
The organic solvents are halogenated hydrocarbons such as methylene
chloride, ethers, such as diethyl ether, tetrahydrofuran or
1,4-dioxane, or aprotic solvents, such as dimethylformamide,
dimethyl sulfoxide or acetonitrile.
The reaction temperature is selected from the range between about
0.degree. C. and around the boiling point of the solvent.
Compound (23) is prepared by cyclizing compound (22) in water, an
organic solvent, or a mixture thereof. The reaction can be carried
out in the presence or absence of a base.
The bases are inorganic bases, such as alkali metal hydroxide (e.g.
sodium hydroxide, potassium hydroxide), alkali metal alkoxide (e.g.
sodium methoxide) or alkali metal carbonates (e.g. potassium
carbonate), or organic bases, such as tertiary amines (e.g.
triethylamine or diisopropylethylamine) or pyridines (e.g.
4-dimethylaminopyridine, pyridine).
The organic solvents are halogenated hydrocarbons such as methylene
chloride, ethers, such as diethyl ether, tetrahydrofuran or
1,4-dioxane, aprotic solvents, such as dimethylformamide or
acetonitrile, or alcohols, such as methanol, ethanol or
2-propanol.
The reaction temperature is selected from the range between room
temperature and around the boiling point of the solvent.
Compound (24) is prepared by dehydrating compound (23) in an
organic solvent.
The dehydration agents are diphosphorus pentaoxide, dicyclohexyl
carbodiimide, etc.
The organic solvents are halogenated hydrocarbons such as methylene
chloride, or aprotic solvents, such as dimethylformamide or
acetonitrile.
The reaction temperature is selected from the range between about
room temperature and around the boiling point of the solvent.
When X is NH, compound (25) is prepared by reacting compound (24)
and guanidine in the presence or absence of a base in an organic
solvent or without any solvent.
The bases are inorganic bases, such as metal alkoxide (e.g. sodium
methoxide), alkali metal hydroxide (e.g. sodium hydroxide), or
alkali metal carbonates (e.g. potassium carbonate), or organic
bases, such as tertiary amines (e.g. triethylamine,
diisopropylethylamine) or pyridines (e.g. 4-dimethylaminopyridine,
pyridine).
The organic solvents are for example, alcohols, such as ethanol or
butanol, ethers, such as tetrahydrofuran or 1,4-dioxane, toluene,
or aprotic solvents, such as dimethylformamide or dimethyl
sulfoxide.
When X is oxygen atom, compound (25) is prepared by reacting
compound (24) and urea in the presence or absence of a base in an
organic solvent or without any solvent.
The bases are inorganic bases, such as metal alkoxide (e.g. sodium
methoxide), alkali metal hydroxide (e.g. sodium hydroxide), or
alkali metal carbonates (e.g. potassium carbonate), or organic
bases such as tertiary amines (e.g. triethylamine) or pyridines
(e.g. dimethylaminopyridine, pyridine).
The organic solvents are for example, alcohols, such as ethanol or
butanol, ethers such as tetrahydrofuran or 1,4-dioxane, toluene, or
aprotic solvents, such as dimethylformamide or dimethyl
sulfoxide.
Compound (25) is also prepared by reacting compound (24) and
benzoyl isocyanate in the presence or absence of a base in an
organic solvent, and then by cyclizing the reactant in the presence
of a base in water, an organic solvent, or a mixture thereof.
The bases used in the reaction with the isocyanate are alkali metal
carbonates such as, potassium carbonate, or organic bases, such as
tertiary amines (e.g. triethylamine, diisopropylethylamine) or
pyridines (e.g. 4-dimethylaminopyridine, pyridine).
The organic solvents are for example, halogenated hydrocarbons such
as methylene chloride, ethers, such as diethyl ether or
tetrahydrofuran, or aprotic solvents, such as dimethylformamide or
dimethyl sulfoxide.
The reaction temperature is selected from the range between about
0.degree. C. and around the boiling point of the solvent.
The organic solvents used in the above cyclizing reaction are for
example, alcohols, such as ethanol or 2-propanol, ethers such as
tetrahydrofuran, or aprotic solvents, such as dimethylformamide or
dimethyl sulfoxide.
The bases are for example, inorganic bases, such as alkali metal
alkoxides (e.g. sodium methoxide), alkali metal hydroxide (e.g.
sodium hydroxide) or ammonia, or organic bases, such as tertiary
amines (e.g. triethylamine) or pyridines (e.g.
4-dimethylaminopyridine, pyridine).
The reaction temperature is selected from the range between around
room temperature and around the boiling point of the solvent.
When X is sulfur atom, compound (25) is prepared by reacting
compound (24) and thiourea in the presence or absence of a base in
an organic solvent or without any solvent.
The bases are metal alkoxides (e.g. sodium methoxide), alkali metal
hydroxides such as sodium hydroxide, alkali metal carbonates such
as potassium carbonate, tertiary amines such as triethylamine, or
pyridines such as 4-dimethylaminopyridine or pyridine.
The organic solvents are alcohols, such as ethanol or butanol,
ethers, such as tetrahydrofuran or 1,4-dioxane, toluene, or aprotic
solvents, such as dimethylformamide or dimethyl sulfoxide.
Compound (25) is also prepared by reacting compound (24) and
benzoyl isothiocyanate in the presence or absence of a base in an
organic solvent, and then by cyclizing the reactant in the presence
of a base in water, an organic solvent, or a mixture thereof.
The bases used in the reaction with the isothiocyanate are alkali
metal carbonates such potassium carbonate, tertiary amines, such as
triethylamine or diisopropylethylamine, or pyridines, such as
4-dimethylaminopyridine or pyridine.
The organic solvents are halogenated hydrocarbons such as methylene
chloride, ethers, such as diethyl ether or tetrahydrofuran, or
aprotic solvents, such as dimethylformamide or dimethyl
sulfoxide.
The reaction temperature is selected from the range between about
0.degree. C. and around the boiling point of the solvent.
The organic solvents used in the above cyclizing reaction are for
example, alcohols such as ethanol or 2-propanol, ethers such as
tetrahydrofuran, or aprotic solvents, such as dimethylformamide or
dimethyl sulfoxide.
The bases are inorganic bases such as metal alkoxides (e.g. sodium
methoxide), alkali metal hydroxide (e.g. sodium hydroxide) or
ammonia, or organic bases such as tertiary amines (e.g.
triethylamine) or pyridines (e.g. 4-dimethylaminopyridine or
pyridine).
The reaction temperature is selected from the range between around
the room temperature and around boiling point of the solvent.
Compound (26) is prepared by reacting compound (25) with R.sup.1 Y
(wherein Y means leaving group such as halogen atom e.g. chlorine
atom, bromine atom) in the presence of a base in an organic
solvent. In this reaction NH.sub.2 or OH group on compound (25) may
be protected or deprotected, if necessary.
The bases are for example, alkali metal hydrogen carbonates such as
sodium hydrogen carbonate, alkali metal carbonates such as sodium
carbonate, tertiary amines such as triethylamine, or pyridines such
as dimethylaminopyridine or pyridine.
The organic solvents are for example, halogenated hydrocarbon such
as methylene chloride, ethers, such as diethyl ether or
tetrahydrofuran, or aprotic solvents such as dimethylformamide.
The reaction temperature is selected from the range between about
0.degree. C. and around the boiling point of the solvent.
When X is NH, compound (26) can be also prepared by reacting
compound (24) and compound (27) in the presence or absence of a
base in an organic solvent or without any solvent.
The bases are metal alkoxides such as sodium methoxide, alkali
metal hydroxides, such as sodium hydroxide, alkali metal
carbonates, such as sodium carbonate, tertiary amines such as
triethylamine, or pyridines, such as 4-dimethylaminopyridine or
pyridine.
The organic solvents are for example, alcohols, such as ethanol or
butanol, ethers, such as tetrahydrofuran or dioxane, toluene, or
aprotic solvents, such as dimethylformamide or dimethyl
sulfoxide.
The compound (I) of the present invention and an intermediate for
preparing it can be purified by the conventional method for
example, column chromatography, recrystallization, etc.
The solvents for the recrystallization are for example, alcohols,
such as methanol, ethanol or 2-propanol, ethers such as diethyl
ether, esters such as ethyl acetate, aromatic hydrocarbon, such as
benzene or toluene, ketones such as acetone, hydrocarbons such as
hexane, or aprotic solvents, such as dimethylformamide or
acetonitrile, or a mixture thereof.
Furthermore, on carrying the above reaction, the protection or
deprotection techniques can be employed, if necessary. The
protection or deprotection techniques are described in detail in
"Protecting group in Organic Synthesis, by T. W. Greene and P. G.
M. Wuts (1990)".
When the compound (I) of the present invention has an asymmetric
carbon atom, optical isomers exist and therefore, a mixture thereof
and an isolated optical isomer are included in the compound (I) of
the present invention.
The compound (I) of the present invention can be orally or
parenterally administered as an interferon inducer. A compound such
that was metabolized in vivo into the compound (I) of the present
invention, or its equivalent compound, so-called "pro-drug" should
be included in the compound of the present invention.
The compound (I) of the present invention is generally administered
in the form of a preparation together with a pharmaceutical
carrier. Said pharmaceutical carriers are selected in accordance to
the form of the preparation, but include for example, starch,
lactose, hydroxypropylcellulose, hydroxypropylmethylcellulose,
polyvinylpyrrolidone, aluminum stearate, magnesium stearate,
etc.
In regards to oral administration, the preparation is administered
in the conventional administration form, for example tablets,
capsules, syrups, or suspensions.
In regard to parenteral administration, the compound is prepared
into solutions, emulsions, suspensions etc., and administered in
the form of injections, or in the form of suppositories,
transdermal formulations or propellants.
Furthermore, the compound may be administered in the form of
sustained release preparations.
Such preparations as mentioned above are prepared by admixing known
carriers, fillers, binders or stabilizing agents with an active
ingredient by a conventional method.
In case of preparing injections, buffer agents, solubilizing
agents, tonicity agents, etc. may be added to them.
Dose and number of administration vary with a disease to be
treated, situation of a patient in question, age, weight, sex, rout
of administration and a kind of preparations. When the preparation
is orally administered, an active ingredient is administered
generally about 1-1000 mg per day, preferably about 10-500 mg, once
or divided into several times. In case of injections, an active
ingredient is administered generally about 0.1-500 mg per day,
preferably about 3-100 mg, once or divided into several times.
The interferon inducer of the present invention can be used as
therapeutic or prophylactic agents such as antivirus agents,
anticancer agents or agents for anti immunologic disease. The route
of administration is oral or parenteral as mentioned above.
EXAMPLE
The examples and reference examples are illustrated as follows.
However, the scope of the present invention should not be limited
to these examples.
Example 1
6-Amino-9-benzyl-8-hydroxy-2-methylthiopurine
##STR8##
6-Amino-9-benzyl-8-bromo-2-methylthiopurine (10 mg, 0.026 mmol) in
concentrated hydrochloric acid (10 ml) was refluxed for 4 hours
under heating. The reaction mixture was made basic with 28% aqueous
ammonia, and the resulting crystals were filtered, washed with
water and dried to give the subject compound (8 mg, yield 96%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 9.60 (1H, br s), 7.31 (5H, m),
6.53 (2H, br s), 4.88 (2H, s), 2.42 (3H, s).
Example 2
6-Amino-9-benzyl-2-ethylthio-8-hydroxypurine
##STR9##
6-Amino-9-benzyl-8-bromo-2-ethylthiopurine (25 mg, 0.069 mmol) in
concentrated hydrochloric acid (25 ml) was refluxed for 4 hours
under heating. The reaction mixture was made basic with 28% aqueous
ammonia, and the resulting crystals were filtered, washed with
water and dried to give the subject compound (6 mg, yield 29%).
.sup.1 H-NMR (DMSO-d6) .delta.: 10.09 (1H, br s), 7.31 (5H, m),
6.51 (2H, br s), 4.88 (2H, s), 2.97 (2H, q, J=7.3 Hz), 1.25 (3H, t,
J=7.3 Hz).
Example 3
6-Amino-9-benzyl-8-hydroxy-2-(propylthio)purine
##STR10##
6-Amino-9-benzyl-8-bromo-2-(propylthio)purine (33 mg, 0.087 mmol)
in concentrated hydrochloric acid (35 ml) was refluxed for 2 hours
under heating. The reaction mixture was made basic with 28% aqueous
ammonia, and the resulting crystals were filtered, washed with
water and dried to give the subject compound (24 mg, yield
87%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.19 (1H, br s), 7.31 (5H,
m), 6.55 (2H, br s), 4.87 (2H, s), 2.98 (2H, t, J=6.9 Hz), 1.61
(2H, m), 0.94 (3H, t, J=7.2 Hz).
Example 4
6-Amino-9-benzyl-8-hydroxy-2-(isopropylthio)purine
##STR11##
6-Amino-9-benzyl-8-bromo-2-(isopropylthio)purine (15 mg, 0.040
mmol) in concentrated hydrochloric acid (20 ml) was refluxed for 2
hours under heating. The reaction mixture was made basic with 28%
aqueous ammonia, and the resulting crystals were filtered, washed
with water and dried to give the subject compound (10 mg, yield
79%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.09 (1H, s), 7.32 (5H, m),
6.50 (2H, br s), 4.87 (2H, s), 3.78 (1H, m), 1.30 (6H, d, J=6.9
Hz).
Example 5
6-Amino-9-benzyl-2-(butylthio)-8-hydroxypurine
##STR12##
6-Amino-9-benzyl-8-bromo-2-(butylthio)purine (23 mg, 0.059 mmol) in
concentrated hydrochloric acid (10 ml) was refluxed for 5 hours
under heating. The reaction mixture was made basic with 28% aqueous
ammonia, and the resulting crystals were filtered, washed with
water and dried to give the subject compound (14 mg, yield
99%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.05 (1H, br s), 7.30 (5H,
m), 6.50 (2H, br s), 4.88 (2H, s), 3.00 (2H, t, J=7.0 Hz), 1.58
(2H, m), 1.35 (2H, m), 0.86 (3H, t, J=7.2 Hz).
Example 6
6-Amino-9-benzyl-8-hydroxy-2-(isobutylthio)purine
##STR13##
6-Amino-9-benzyl-8-bromo-2-(isobutylthio)purine (21 mg, 0.053 mmol)
in concentrated hydrochloric acid (20 ml) was refluxed for 5 hours
under heating. The reaction mixture was made basic with 28% aqueous
ammonia, and the resulting crystals were filtered, washed with
water and dried to give the subject compound (16 mg, yield
91%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.10 (1H, s), 7.26-7.35 (5H,
m), 6.51 (2H, br s), 4.87 (2H, s), 2.93 (2H, d, J=6.6 Hz), 1.83(1H,
m), 0.93 (6H, d, J=6.6 Hz).
Example 7
6-Amino-9-benzyl-8-hydroxy-2-(sec-butylthio)purine
##STR14##
6-Amino-9-benzyl-8-bromo-2-(sec-butylthio)purine (39 mg, 0.092
mmol) in concentrated hydrochloric acid (20 ml) was refluxed for 2
hours under heating. The reaction mixture was made basic with 28%
aqueous ammonia, and the resulting crystals were filtered, washed
with water and dried to give the subject compound (12 mg, yield
40%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.09 (1H, br s), 7.24-7.35
(5H, m), 6.50 (2H, br s), 4.87 (2H, s), 3.65 (1H, m), 1.61 (2H, m),
1.28 (3H, d, J=7.0 Hz), 0.93 (3H, t, J=7.3 Hz).
Example 8
6-Amino-9-benzyl-hydroxy-2-(pentylthio)purine
##STR15##
6-Amino-9-benzyl-8-bromo-2-(pentylthio)purine (39 mg, 0.096 mmol)
in concentrated hydrochloric acid (35 ml) was refluxed for 2.5
hours under heating. The reaction mixture was made basic with 28%
aqueous ammonia, and the resulting crystals were filtered, washed
with water and dried to give the subject compound (30 mg, yield
91%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.05 (1H, br s), 7.30 (5H,
m), 6.50 (2H, br s), 4.88 (2H, s), 2.99 (2H, t, J=7.3 Hz), 1.59
(2H, m), 1.30 (4H, m), 0.84 (3H, t, J=7.3 Hz).
Example 9
6-Amino-9-benzyl-8-hydroxy-2-13-methylbutylthio) purine
##STR16##
6-Amino-9-benzyl-8-bromo-2-(3-methylbutylthio)purine (11 mg, 0.027
mmol) in concentrated hydrochloric acid (20 ml) was refluxed for 3
hours under heating. The reaction mixture was made basic with 28%
aqueous ammonia, and the resulting crystals were filtered, washed
with water and dried to give the subject compound (7 mg, yield
75%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.10 (1H, br s), 7.30 (5H,
m), 6.50 (2H, br s), 4.88 (2H, s), 3.00 (2H, t, J=7.6 Hz) 1.63 (1H,
m), 1.51 (2H, m), 0.86 (6H, t, J=6.2 Hz).
Example 10
6-Amino-9-benzyl-8-hydroxy-2-(2-methylbutylthio)purine
##STR17##
6-Amino-9-benzyl-8-bromo-2-(2-methylbutylthio)purine (29 mg, 0.071
mmol) in concentrated hydrochloric acid (20 ml) was refluxed for 3
hours under heating. The reaction mixture was made basic with 28%
aqueous ammonia, and the resulting crystals were filtered, washed
with water and dried to give the subject compound (6 mg, yield
25%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.08 (1H, s), 7.30 (5H, m),
6.50 (2H, br s), 4.88 (2H, s), 3.08 (1H, q, J=6.6 Hz), 2.86 (1H,
m), 1.62 (1H, m), 1.43 (1H, m), 1.15 (1H, m), 0.91 (3H, d, J=6.6
Hz), 0.86 (3H, t, J=6.2 Hz).
Example 11
6-Amino-9-benzyl-2-cyclohexylthio-8-hydroxypurine
##STR18##
6-Amino-9-benzyl-8-bromo-2-cyclohexylthiopurine (20 mg, 0.048 mmol)
in concentrated hydrochloric acid (10 ml) was refluxed for 6 hours
under heating. The reaction mixture was made basic with 28% aqueous
ammonia, and the resulting crystals were filtered, washed with
water and dried to give the subject compound (12 mg, yield
70%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.09 (1H, br s), 7.31 (5H,
m), 6.49 (2H, br s), 4.87 (2H, s), 3.62 (1H, m), 2.00 (2H, m), 1.68
(2H, m), 1.62-1.56 (1H, m), 1.35 (5H, m).
Example 12
6-Amino-9-benzyl-8-hydroxy-2-phenylthiopurine
##STR19##
6-Amino-9-benzyl-8-bromo-2-phenylthiopurine (31 mg, 0.075 mmol) in
concentrated hydrochloric acid (20 ml) was refluxed for 12 hours
under heating. The reaction mixture was made basic with 28% aqueous
ammonia, and the resulting crystals were filtered, washed with
water and dried to give the subject compound (11 mg, yield
42%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 9.50 (1H, br s), 7.55 (2H, m),
7.46 (3H, m), 7.28 (3H, m), 7.13 (2H, m), 6.55 (2H, br s), 4.67
(2H, s).
Example 13
6-Amino-9-benzyl-8-hydroxy-2-(p-tolylthio)purine
##STR20##
6-Amino-9-benzyl-8-bromo-2-(p-tolylthio)purine (15 mg, 0.035 mmol)
in concentrated hydrochloric acid (20 ml) was refluxed for 7.5
hours under heating. The reaction mixture was made basic with 28%
aqueous ammonia, and the resulting crystals were filtered, washed
with water and dried to give the subject compound (5 mg, yield
39%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 7.44 (2H, d, J=7.9 Hz), 7.27
(5H, m), 7.13 (2H, m), 6.51 (2H, br s), 4.67 (2H, s), 2.35 (3H,
s).
Example 14
6-Amino-9-benzyl-8-hydroxy-2-(2-naphthylthio)purine
##STR21##
6-Amino-9-benzyl-8-bromo-2-(2-naphthylthio)purine (33 mg, 0.043
mmol) in a mixture of concentrated hydrochloric acid (20 ml) and
dimethyl sulfoxide (7 ml) was refluxed for 6 hours under heating.
The reaction mixture was made basic with 28% aqueous ammonia, and
the resulting crystals were filtered, washed with water and the
crude product was purified by thin-layer chromatography to give the
subject compound (6 mg, yield 35%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.14 (1H, br s), 8.30 (1H, d,
J=8.6 Hz), 7.98-8.07 (2H, m), 7.67-7.77 (3H, m), 7.12-7.20 (3H, m),
6.69 (2H, d, J=6.9 Hz), 6.59 (2H, br s), 4.58 (2H, s).
Example 15
6-Amino-9-benzyl-2-benzylthio-8-hydroxypurine
##STR22##
6-Amino-9-benzyl-2-benzylthio-8-bromopurine (18 mg, 0.042 mmol) in
concentrated hydrochloric acid (10 ml) was refluxed for 9 hours
under heating. The reaction mixture was made basic with 28% aqueous
ammonia, and the resulting crystals were filtered, washed with
water and dried to give the subject compound (8 mg, yield 52%).
.sup.1 H-NMR (DMSO-ds) .delta.: 10.12 (1H, br s), 7.19-7.34 (10H,
m), 6.58 (2H, br s), 4.91 (2H, s), 4.29 (2H, s).
Example 16
6-Amino-9-benzyl-8-hydroxy-2-methoxypurine
##STR23##
6-Amino-9-benzyl-2,8-dimethoxypurine (53 mg, 0.186 mmol) in
concentrated hydrochloric acid (10 ml) was stirred for 3 hours at
room temperature. The reaction mixture was made basic with 28%
aqueous ammonia, and the resulting crystals were filtered and
washed with water to give the subject compound (38 mg, yield
75%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 9.95 (1H, br s), 7.35-7.22
(5H, m), 6.50 (2H, br s), 4.86 (2H, s), 3.76 (3H, s).
Example 17
6-Amino-9-benzyl-2-ethoxy-8-hydroxypurine
##STR24##
6-Amino-9-benzyl-2-ethoxy-8-methoxypurine (18 mg, 0.06 mmol) in
concentrated hydrochloric acid (5 ml) was stirred for 3 hours at
room temperature. The reaction mixture was made basic with 28%
aqueous ammonia, and the resulting crystals were filtered and
washed with water to give the subject compound (11 mg, yield
64%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 9.96 (1H, br s), 7.35-7.23
(5H, m), 6.45 (2H, br s), 4.85 (2H, s), 4.19 (2H, q, J=7.1 Hz),
1.25 (3H, t, J=7.1 Hz).
Example 18
6-Amino-9-benzyl-8-hydroxy-2-propoxypurine
##STR25##
6-Amino-9-benzyl-8-methoxy-2-propoxypurine (75 mg, 0.24 mmol) in
concentrated hydrochloric acid (15 ml) was stirred for 3 hours at
room temperature. The reaction mixture was made basic with 28%
aqueous ammonia, and the resulting crystals were filtered and
washed with water to give the subject compound (59 mg, yield
83%).
.sup.1 H-NMR (DMSO-d) .delta.: 9.96 (1H, br s), 7.35-7.22 (5H, m),
6.45 (2H, br s), 4.86 (2H, s), 4.10 (2H, t, J=6.8 Hz), 1.65 (2H,
m), 0.93 (3H, t, J=7.3 Hz).
Example 19
6-Amino-9-benzyl-2-butoxy-8-hydroxypurine
##STR26##
6-Amino-9-benzyl-2-butoxy-8-methoxypurine (20 mg, 0.061 mmol) in
concentrated hydrochloric acid (5 ml) was stirred for 3 hours at
room temperature. The reaction mixture was made basic with 28%
aqueous ammonia. The resulting crystals were filtered, washed with
water, and the crude product was purified by silica gel
chromatography (3% methanol/chloroform) to give the subject
compound (13 mg, yield 68%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 9.95 (1H, br s), 7.35-7.23
(5H, m), 6.45 (2H, br s), 4.86 (2H, s), 4.13 (2H, t, J=6.4 Hz),
1.62 (2H, m), 1.37 (2H, m), 0.90 (3H, t, J=7.3 Hz).
Example 20
6-Amino-9-benzyl-8-hydroxy-2-pentoxypurine
##STR27##
6-Amino-9-benzyl-8-methoxy-2-pentoxypurine (40 mg, 0.117 mmol) in
concentrated hydrochloric acid (20 ml) was stirred for 12 hours at
room temperature. The reaction mixture was made basic with 28%
aqueous ammonia. The resulting crystals were filtered and washed
with water to give the subject compound (33 mg, yield 86%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 9.97 (1H, br s), 7.35-7.24
(5H, m), 6.44 (2H, br s), 4.85 (2H, s), 4.13 (2H, t, J=6.6 Hz),
1.62 (2H, m), 1.32 (4H, m), 0.88 (3H, t, J=6.4 Hz).
Example 21
6-Amino-9-benzyl-8-hydroxy-2-methylaminopurine
##STR28##
6-Amino-9-benzyl-8-bromo-2-methylaminopurine (55 mg, 0.17 mmol) in
concentrated hydrochloric acid (30 ml) was refluxed for 5 hours
under heating. The reaction mixture was made basic with 28% aqueous
ammonia. The resulting crystals were filtered, washed with water
and dried to give the subject compound (42 mg, yield 94%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 9.67 (1H, br s), 7.31-7.24
(5H, m), 6.19 (1H, q, J=4.8 Hz), 6.06 (2H, br s), 4.81 (2H, s),
2.69 (3H, d, J=4.8 Hz).
Example 22
6-Amino-9-benzyl-2-ethylamino-8-hydroxypurine
##STR29##
6-Amino-9-benzyl-8-bromo-2-ethylaminopurine (55 mg, 0.16 mmol) in
concentrated hydrochloric acid (30 ml) was refluxed for 5 hours
under heating. The reaction mixture was made basic with 28% aqueous
ammonia. The resulting crystals were filtered, washed with water
and dried to give the subject compound (45 mg yield:
quantitatively).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 9.65 (1H, br s), 7.34-7.24
(5H, m), 6.18 (1H, t, J=5.55 Hz), 6.01 (2H, br s), 4.81 (2H, s),
3.19 (2H, m), 1.06 (3H, t, J=7.1 Hz).
Example 23
6-Amino-9-benzyl-8-hydroxy-2-propylaminopurine
##STR30##
6-Amino-9-benzyl-8-bromo-2-propylaminopurine (86 mg, 0.24 mmol) in
concentrated hydrochloric acid (30 ml) was refluxed for 5 hours
under heating. The reaction mixture was made basic with 28% aqueous
ammonia. The resulting crystals were filtered, washed with water
and dried to give the subject compound (69 mg, yield 97%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 9.64 (1H, br s), 7.34-7.24
(5H, m), 6.22 (1H, t, J=5.5 Hz), 6.00 (2H, br s), 4.80 (2H, s),
3.12 (2H, m), 1.46 (2H, m), 0.85 (3H, t, J=7.5 Hz).
Example 24
6-Amino-9-benzyl-2-butylamino-8-hydroxypurine
##STR31##
6-Amino-9-benzyl-8-bromo-2-butylaminopurine (78 mg, 0.21 mmol) in
concentrated hydrochloric acid (30 ml) was refluxed for 5 hours
under heating. The reaction mixture was made basic with 28% aqueous
ammonia. The resulting crystals were filtered, washed with water
and dried to give the subject compound (54 mg, yield 83%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 9.64 (1H, br s), 7.29-7.24
(5H, m), 6.19 (1H, t, J=6.2 Hz), 6.00 (2H, br s), 4.80 (2H, s),
3.15 (2H, m), 1.43 (2H, m), 1.28 (2H, m), 0.87 (3H, t, J=7.3
Hz).
Example 25
6-Amino-9-benzyl-8-hydroxy-2-pentylaminopurine
##STR32##
6-Amino-9-benzyl-8-bromo-2-pentylaminopurine (74 mg, 0.19 mmol) in
concentrated hydrochloric acid (20 ml) was refluxed for 5 hours
under heating. The reaction mixture was made basic with 28% aqueous
ammonia. The resulting crystals were filtered, washed with water
and dried to give the subject compound (36 mg, yield 58%).
.sup.1 H-NMR (DMSO-ds) .delta.: 9.63 (1H, br s), 7.30-7.24 (5H, m),
6.19 (1H, t, J=5.3 Hz), 5.99 (2H, br s), 4.80 (2H, s), 3.19-3.11
(2H, m), 1.48-1.43 (2H, m), 1.27-1.24 (4H, m), 0.85 (3H, t, J=7.0
Hz).
Example 26
6-Amino-9-benzyl-8-hydroxy-2-(isopropylamino)purine
##STR33##
6-Amino-9-benzyl-8-bromo-2-(isopropylamino)purine (68 mg, 0.19
mmol) in concentrated hydrochloric acid (20 ml) was refluxed for 5
hours under heating. The reaction mixture was made basic with 28%
aqueous ammonia. The resulting crystals were filtered, washed with
water and dried to give the subject compound (50 mg, yield
89%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 9.64 (1H, br s), 7.34-7.21
(5H, m), 5.99 (2H, br s), 5.98 (1H, d, J=8.2 Hz), 4.80 (2H, s),
4.00-3.90 (1H, m), 1.08 (6H, d, J=6.4 Hz).
Example 27
6-Amino-9-benzyl-8-hydroxy-2-(isobutylamino)purine
##STR34##
6-Amino-9-benzyl-8-bromo-2-(isobutylamino)purine (55 mg, 0.19 mmol)
in concentrated hydrochloric acid (20 ml) was refluxed for 5 hours
under heating. The reaction mixture was made basic with 28% aqueous
ammonia. The resulting crystals were filtered, washed with water
and dried to give the subject compound (30 mg, yield 52%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 9.63 (1H, br s), 7.30-7.22
(5H, m), 6.24 (1H, t, J=6.0 Hz), 5.99 (2H, br s), 4.80 (2H, s),
2.99 (2H, dd, J=6.0, 6.0 Hz), 1.84-1.75 (1H, m), 0.84 (6H, d, J=6.8
Hz).
Example 28
6-Amino-9-benzyl-8-hydroxy-2-(sec-butylamino)purine
##STR35##
6-Amino-9-benzyl-8-bromo-2-(sec-butylamino)purine (50 mg, 0.13
mmol) in concentrated hydrochloric acid (20 ml) was refluxed for 5
hours under heating. The reaction mixture was made basic with 28%
aqueous ammonia. The resulting crystals were filtered, washed with
water and dried to give the subject compound (23 mg, yield
55%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 9.63 (1H, br s), 7.31-7.24
(5H, m), 5.97 (2H, br s), 5.95 (1H, d, J=8.6 Hz), 4.80 (2H, s),
3.82-3.74 (1H, m), 1.51-1.34 (2H, m), 1.04 (3H, d, J=6.4 Hz), 0.83
(3H, t, J=7.3 Hz).
Example 29
6-Amino-9-benzyl-2-(2.2-dimethylpropyl)amino-8-hydroxypurine
##STR36##
6-Amino-9-benzyl-8-bromo-2-(2,2-dimethylpropyl)aminopurine (70 mg,
0.18 mmol) in concentrated hydrochloric acid (20 ml) was refluxed
for 5 hours under heating. The reaction mixture was made basic with
28% aqueous ammonia. The resulting crystals were filtered, washed
with water and dried to give the subject compound (23 mg, yield
39%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 9.63 (1H, br s), 7.28-7.21
(5H, m), 6.04 (1H, t, J=6.2 Hz), 5.97 (2H, br s), 4.79 (2H, s),
3.06 (2H, d, J=6.4 Hz), 0.83 (9H, s).
Example 30
6-Amino-9-benzyl-2-benzylamino-8-hydroxypurine
##STR37##
6-Amino-9-benzyl-2-benzylamino-8-bromopurine (37 mg, 0.09 mmol) in
concentrated hydrochloric acid (50 ml) was refluxed for 5 hours
under heating. The reaction mixture was made basic with 28% aqueous
ammonia. The resulting crystals were filtered, washed with water
and dried to give the subject compound (7 mg, yield 23%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 9.75 (1H, br s), 7.31-7.15
(10H, m), 6.83 (1H, t, J=6.4 Hz), 6.10 (2H, br s), 4.78 (2H, s),
4.40 (2H, d, J=6.4 Hz).
Example 31
6-Amino-9-benzyl-2-cyclohexylamino-8-hydroxypurine
##STR38##
6-Amino-9-benzyl-8-bromo-2-cyclohexylaminopurine (82 mg, 0.20 mmol)
in concentrated hydrochloric acid (30 ml) and methanol (20 ml) were
refluxed for 5 hours under heating. After removal of methanol, the
reaction mixture was made basic with 28% aqueous ammonia. The
resulting crystals were filtered, washed with water and dried to
give the subject compound (7 mg, yield 23%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 9.62 (1H, br s), 7.28 (5H, m),
5.96 (3H, br s), 4.78 (2H, s), 3.58 (1H, m), 1.80 (2H, m), 1.65
(2H, m), 1.56 (1H, m), 1.27-1.06 (5H, m).
Example 32
6-Amino-2-anilino-9-benzyl-8-hydroxypurine
##STR39##
6-Amino-2-anilino-9-benzyl-8-bromopurine (80 mg, 0.20 mmol) in
concentrated hydrochloric acid (200 ml) and methanol (50 ml) were
refluxed for 5 hours under heating. The reaction mixture was
condensed in vacuo, 28% aqueous ammonia was added to the residue.
The resulting solid was filtered, washed with water and dried to
give the subject compound (67 mg yield: quantitatively).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.00 (1H, Br s), 8.22 (1H, d,
J=8.8 Hz), 7.80 (1H, d, J=2.2 Hz), 7.54 (1H, s), 7.47 (1H, dd,
J=8.8, 2.2 Hz), 7.35-7.26 (7H, m), 6.45 (2H, br s), 4.89 (2H,
s).
Example 33
6-Amino-9-benzyl-2-dimethylamino-8-hydroxypurine
##STR40##
6-Amino-9-benzyl-2-dimethylamino-8-bromopurine (51 mg, 0.15 mmol)
in concentrated hydrochloric acid (30 ml) and methanol (10 ml) were
refluxed for 5 hours under heating. After removal of methanol, the
reaction mixture was made basic with 28% aqueous ammonia. The
resulting crystals were filtered, washed with water and dried to
give the subject compound (38 mg, yield 91%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 9.67 (1H, br s), 7.36-7.24
(5H, m), 6.08 (2H, br s), 4.82 (2H, s), 3.01 (6H, s).
Example 34
6-Amino-9-benzyl-2-benzylmethylamino-8-hydroxypurine
##STR41##
6-Amino-9-benzyl-2-benzylmethylamino-8-bromopurine (85 mg, 0.20
mmol) in concentrated hydrochloric acid (30 ml) and methanol (20
ml) were refluxed for 5 hours under heating. The reaction mixture
was made basic with 28% aqueous ammonia, and the resulting crystals
were filtered, washed with water and dried to give the subject
compound (56 mg, yield 77%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 9.70 (1H, br s), 7.29-7.19
(10H, m), 6.12 (2H, br s), 4.81 (2H, s), 4.77 (2H, s), 2.99 (3H,
s).
Example 35
6-Amino-9-benzyl-8-hydroxy-2-(2-phthalimidoethyl)thiopurine
##STR42##
6-Amino-9-benzyl-8-hydroxy-2-mercaptopurine (120 mg, 0.44 mmol) was
suspended in dimethylformamide (10 ml). To the suspension were
added potassium carbonate (60 mg, 0.43 mmol) and 2-phthalimidoethyl
bromide (112 mg, 0.44 mmol) in order. The mixture was stirred at
room temperature for 2 hours. The solvent was removed in vacuo and
to the residue was added saturated brine. The mixture was extracted
with ethyl acetate, the organic layer was dried on magnesium
sulfate, and the solvent was removed in vacuo. Methanol was added
to the residue, and the resulting crystals were taken by filtration
to give the subject compound (107 mg, yield 54%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.19 (1H, br s), 7.83 (4H,
m), 7.34 (5H, m), 6.52 (2H, br s), 4.96 (2H, s), 3.95 (2H, t, J=6.6
Hz), 3.32 (2H, t, J=6.6 Hz).
Example 36
6-Amino-9-benzyl-8-hydroxy-2-(3-phthalimidopropylthio)purine
##STR43##
6-Amino-9-benzyl-8-hydroxy-2-mercaptopurine (110 mg, 0.40 mmol) was
suspended in dimethylformamide (10 ml). To the suspension were
added potassium carbonate (50 mg, 0.40 mmol) and 2-phthalimidoethyl
bromide (108 mg, 0.40 mmol) in order. The mixture was stirred at
room temperature for 2 hours. The solvent was removed in vacuo,
water and methanol were added to the residue, and the resulting
crystals were taken by filtration to give the subject compound (138
mg, yield 75%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.09 (1H, s), 7.82 (4H, m),
7.24 (5H, m), 6.50 (2H, br s), 4.82 (2H, s), 3.67 (2H, t, J=6.2
Hz), 3.03 (2H, t, J=6.2 Hz), 1.96 (2H, m).
Example 37
6-Amino-9-benzyl-8-hydroxy-2-(4-phthalimidobutylthio)purine
##STR44##
6-Amino-9-benzyl-8-hydroxy-2-mercaptopurine (120 mg, 0.44 mmol) was
suspended in dimethylformamide (10 ml). To the suspension were
added potassium carbonate (60 mg, 0.43 mmol) and 4-phthalimidobutyl
bromide (113 mg, 0.40 mmol) in order. The mixture was stirred at
room temperature for 2 hours. The solvent was removed in vacuo, and
water and methanol were added to the residue, and the resulting
crystals were taken by filtration to give the subject compound (141
mg, yield 74%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.09 (1H, br s), 7.84 (4H,
m), 7.29 (5H, m), 6.51 (2H, br s), 4.83 (2H, s), 3.56 (2H, t, J=6.3
Hz), 3.03 (2H, t, J=6.9 Hz), 1.67 (4H, m).
Example 38
3-[(6-Amino-9-benzyl-8-hydroxy-2-purinyl)thio]propanol
##STR45##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (200 mg, 0.73
mmol) was suspended in dimethylformamide (100 ml). To the
suspension were added potassium carbonate (150 mg, 1.1 mmol) and
3-bromo-1-propanol (0.1 ml, 1 mmol) in order. The mixture was
stirred at room temperature for 4 hours. The solvent was removed in
vacuo, and the residue was purified by silica gel chromatography
(1% methanol/chloroform) to give the subject compound (149 mg,
yield 62%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.09 (1H, s), 7.31 (5H, m),
6.50 (2H, br s), 4.90 (2H, s), 4.50 (1H, t, J=5.6 Hz), 3.49 (2H,
m), 3.07 (2H, t, J=6.6 Hz), 1.75 (2H, m).
Example 39
6-Amino-9-henzyl-8-hydroxy-2-(methoxycarbonylmethylthio)purine
##STR46##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (200 mg, 0.73
mmol) was suspended in dimethylformamide (80 ml). To the suspension
were added potassium carbonate (150 mg, 1.1 mmol) and methyl
bromoacetate (0.1 ml, 1.1 mmol) in order. The mixture was stirred
at room temperature for 2 hours. The solvent was removed in vacuo,
and the residue was purified by silica gel chromatography (3%
methanol/chloroform) to give the subject compound (173 mg, yield
69%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.12 (1H, s), 7.30 (5H, m),
6.57 (2H, br s), 4.84 (2H, s), 3.91 (2H, m), 3.56 (2H, s).
Example 40
6-Amino-9-benzyl-8-hydroxy-2-[2-(methoxycarbonyl)ethyl]thiopurine
##STR47##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (200 mg, 0.73
mmol) was suspended in dimethylformamide (80 ml). To the suspension
were added potassium carbonate (150 mg, 1.1 mmol) and methyl
3-bromopropionate (0.12 ml, 1.1 mmol) in order. The mixture was
stirred at room temperature for 2 hours. The solvent was removed in
vacuo, and the residue was purified by silica gel chromatography
(3% methanol/chloroform) to give the subject compound (146 mg,
yield 56%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.12 (1H, s), 7.30 (5H, m),
6.57 (2H, br s), 4.84 (2H, s), 3.91 (2H, s), 3.56 (2H, s).
Exanple 41
6-Amino-9-benzyl-8-hydroxy-2-(carboxymethylthio)purine
##STR48##
To a methanol solution (5 ml) containing 500 mg of sodium hydroxide
was added
6-amino-9-benzyl-8-hydroxy-2-(methoxycarbonylmethyl)thiopurine (64
mg, 0.19 mmol). The solution was refluxed under heating,
neutralized with 2N hydrochloric acid, filtered and washed with
water to give the subject compound (32 mg, yield 52%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.44 (1H, s), 7.34 (5H, m),
6.64 (2H, br s), 4.85 (2H, s), 3.82 (2H, s).
Example 42
6-Amino-9-benzyl-8-hydroxy-2-(methoxymethylthio)purine
##STR49##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (134 mg, 0.49
mmol) was suspended in dimethylformamide (65 ml). To the suspension
were added potassium carbonate (100 mg, 0.72 mmol) and chloromethyl
methyl ether (0.056 ml, 0.73 mmol) in order. The mixture was
stirred at room temperature for 2 hours. The solvent was removed in
vacuo, and the residue was purified by silica gel chromatography
(2% methanol/chloroform) to give the subject compound (107 mg,
yield 69%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.15 (1H, s), 7.31 (5H, m),
6.59 (2H, br s), 5.29 (2H, s), 4.89 (2H, s), 3.21 (3H, s).
Example 43
6-Amino-9-benzyl-8-hydroxy-2-(2-ethoxyethyl)thiopurine
##STR50##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (134 mg, 0.49
mmol) was suspended in dimethylformamide (65 ml). To the suspension
were added potassium carbonate (100 mg, 0.72 mmol) and
2-chloroethyl ethyl ether (0.056 ml, 0.73 mmol) in order. The
mixture was stirred at room temperature for 3 hours. The solvent
was removed in vacuo, and the residue was purified by silica gel
chromatography (1% methanol/chloroform) to give the subject
compound (19 mg, yield 11%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.11 (1H, s), 7.30 (5H, m),
6.54 (2H, br s), 4.88 (2H, s), 3.54 (2H, t, J=6.9 Hz), 3.43 (2H, q,
J=7.0 Hz), 3.18 (2H, t, J=6.6 Hz), 1.08 (3H, t, J=6.9 Hz).
Example 44
6-Amino-9-benzyl-8-hydroxy-2-[(2-hydroxyethyl)thio]purine
##STR51##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (134 mg, 0.49
mmol) was suspended in dimethylformamide (65 ml). To the suspension
were added potassium carbonate (100 mg, 0.72 mmol) and
2-bromoethanol (0.052 ml, 0.73 mmol) in order. The mixture was
stirred at room temperature for 5 hours. The solvent was removed in
vacuo, and the residue was purified by silica gel chromatography
(2% methanol/chloroform) to give the subject compound (72 mg, yield
46%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.09 (1H, s), 7.32 (5H, m),
6.52 (2H, br s), 4.87 (3H, s), 3.59 (2H, q, J=5.9 Hz), 3.12 (2H, t,
J=6.6 Hz).
Example 45
[(6-Amino-9-benzyl-8-hydroxy-2-purinyl)thio]acetamide
##STR52##
A 28% ammonia/methanol solution was added to
6-amino-9-benzyl-8-hydroxy-2-(methoxycarbonylmethyl)thiopurine (75
mg, 0.22 mmol). The solution was heated in autoclave for 6 hours
and then the solvent was removed in vacuo. To the residue was added
methanol and the resulting crystals were filtered to give the
subject compound (64 mg, yield 89%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.12 (1H, s), 7.42 (1H, br
s), 7.34 (5H, m), 7.07 (1H, br s), 6.57 (2H, br s), 4.87 (2H, s),
3.70 (2H, s).
Example 46
6-Amino-9-benzyl-2-[(1,3-dioxolan-2-yl)-methyl]thio-8-hydroxypurine
##STR53##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (200 mg, 0.73
mmol) was suspended in dimethylformamide (80 ml). To the suspension
were added potassium carbonate (150 mg, 1.1 mmol) and
2-bromomethyl-1,3-dioxolane (0.11 ml, 1.1 mmol) in order. The
mixture was stirred at room temperature for 3 hours. The solvent
was removed in vacuo, and the residue was purified by silica gel
chromatography (3% methanol/chloroform) to give the subject
compound (73 mg, yield 28%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.12 (1H, s), 7.31 (5H, m),
6.57 (2H, br s), 5.03 (1H, t, J=4.6 Hz), 4.87 (2H, s), 3.92 (2H,
m), 3.78 (2H, m), 3.28 (2H, d, J=5.0 Hz).
Example 47
6-Amino-9-benzyl-8-hydroxy-2-[2-(dimethylamino)ethyl)thiopurine
##STR54##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (200 mg, 0.73
mmol) was suspended in dimethylformamide (80 ml). To the suspension
were added potassium carbonate (300 mg, 2.2 mmol) and
2-dimethylaminoethylchloride (160 mg, 1.1 mmol) in order. The
mixture was stirred at room temperature for 11 hours. The solvent
was removed in vacuo, and the residue was purified by silica gel
chromatography (10% methanol/chloroform) to give the subject
compound (9 mg, yield 4%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.11 (1H, s), 7.29 (5H, m),
6.53 (2H, br s), 4.88 (2H, s), 3.11 (2H, t, J=7.6 Hz), 2.14 (6H,
s).
Example 48
6-Amino-9-benzyl-8-hydroxy-2-[(2-methoxyethyl)thio]purine
##STR55##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (134 mg, 0.49
mmol) was suspended in dimethylformamide (65 ml). To the suspension
were added potassium carbonate (100 mg, 0.72 mmol) and
2-chloroethyl methyl ether (0.067 ml, 0.73 mmol) in order. The
mixture was stirred at room temperature for 8 hours. The solvent
was removed in vacuo, and the residue was purified by silica gel
chromatography (3% methanol/chloroform) to give the subject
compound (20 mg, yield 12%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.11 (1H, s), 7.30 (5H, m),
6.55 (2H, br s), 4.87 (2H, s), 3.50 (2H, t, J=6.6 Hz), 3.22 (3H,
m), 3.21 (2H, t, J=6.6 Hz).
Example 49
6-Amino-9-benzyl-8-hydroxy-2-(formylmethylthio)purine
##STR56##
6-Amino-9-benzyl-2-(1,3-dioxolan-2-yl-methyl)thio-8-hydroxypurine
(44 mg, 0.12 mmol) was dissolved in a mixture of 3.3N hydrochloric
acid (1 ml) and tetrahydrofuran (4 ml). The solution was stirred at
70.degree. C. for 7 hours and then neutralized with aqueous sodium
hydrogen carbonate. After removal of tetrahydrofuran in vacuo, the
resulting crystals were filtered, washed with water and repulped in
methanol to give the subject compound (17 mg, yield 44%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.14 (1H, br s), 9.52 (1H,
s), 7.31 (5H, m), 6.60 (2H, br s), 3.81 (2H, s).
Example 50
6-Amino-9-benzyl-8-hydroxy-2-(2-morpholinoethyl)thiopurine
##STR57##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (134 mg, 0.49
mmol) was suspended in dimethylformamide (65 ml). To the suspension
were added potassium carbonate (100 mg, 0.72 mmol) and
4-(2-chloroethyl)morpholine (136 mg, 0.73 mmol) in order. The
mixture was stirred at room temperature for 8 hours. The solvent
was removed in vacuo, and the residue was purified by silica gel
chromatography (8% methanol/chloroform) to give the subject
compound (34 mg, yield 18%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.12 (1H, s), 7.28 (511, m),
6.53 (2H, br s), 3.51 (4H, m), 3.11 (2H, t, J=7.6 Hz), 2.34 (4H,
m).
Example 51
6-Amino-9-benzyl-2-[2-(1,3-dioxolan-2-yl)ethylthio)-8-hydroxypurine
##STR58##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (200 mg, 0.73
mmol) was suspended in dimethylformamide (80 ml). To the suspension
were added potassium carbonate (150 mg, 1.1 mmol) and
2-(2-bromoethyl)-1,3-dioxolane (0.14 ml, 1.1 mmol) in order. The
mixture was stirred at room temperature for 3 hours. The solvent
was removed in vacuo, and the residue was purified by silica gel
chromatography (3% methanol/chloroform) to give the subject
compound (163 mg, yield 60%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.10 (1H, s), 7.34 (5H, m),
6.52 (2H, br s), 4.89 (1H, t, J=4.3 Hz), 4.87 (2H, s), 3.89 (2H,
m), 3.77 (2H, m), 3.05 (2H, t, J=6.9 Hz), 1.96 (2H, m).
Example 52
6-Amino-9-benzyl-2-(2-formylethylthio)-8-hydroxypurine
##STR59##
6-Amino-9-benzyl-2-[2-(1,3-dioxolan-2-yl)ethyl)thio-8-hydroxypurine
(74 mg, 0.20 mmol) was dissolved in a mixture of 3.3N hydrochloric
acid (1 ml) and tetrahydrofuran (4 ml). The solution was stirred at
70.degree. C. for 7 hours and then neutralized with aqueous sodium
hydrogen carbonate. After removal of tetrahydrofuran in vacuo, the
resulting crystals were filtered, purified by silica gel
chromatography (5% methanol/chloroform) to give the subject
compound (17 mg, yield 44%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.12 (1H, s), 9.64 (1H, s),
7.30 (5H, m), 6.55 (2H, br s), 4.86 (2H, s), 3.02 (2H, t, J=6.9
Hz), 2.82 (2H, t, J=7.0 Hz).
Example 53
6-Amino-9-benzyl-8-hydroxy-2-[(2-carboxyethyllthio]purine Acid
##STR60##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (200 mg, 0.73
mmol) was suspended in dimethylformamide (60 ml). To the suspension
were added potassium carbonate (300 mg, 2.2 mmol) and
3-iodopropionic acid (220 mg, 1.1 mmol) in order. The mixture was
stirred at room temperature for 3 hours. The solvent was removed in
vacuo, and the residue was purified by silica gel chromatography
(20% methanol/chloroform) to give the subject compound (38 mg,
yield 15%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 12.27 (1H, s), 10.12 (5H, m),
7.31 (5H, m), 6.55 (2H, br s), 4.87 (2H, s), 3.17 (2H, t, J=6.6
Hz), 2.62 (2H, t, J=6.6 Hz).
Example 54
6-Amino-9-benzyl-8-hydroxy-2-[(2,2,2-trifluoroethyl)thio]purine
##STR61##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (134 mg, 0.49
mmol) was suspended in dimethylformamide (65 ml). To the suspension
were added potassium carbonate (100 mg, 0.7 mmol) and
2-iodo-1,1,1-trifluoroethane (0.07 ml, 0.73 mmol) in order. The
mixture was stirred at room temperature for 8 hours. The solvent
was removed in vacuo, and the residue was purified by silica gel
chromatography (3% methanol/chloroform) to give the subject
compound (20 mg, yield 12%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.19 (1H, s), 7.31 (5H, m),
6.70 (2H, br s), 4.90 (2H, s), 4.16 (2H, q, J=10.5 Hz).
Example 55
6-Amino-9-benzyl-2-[(2-fluoroethyl)thio]-8-hydroxypurine
##STR62##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (134 mg, 0.49
mmol) was suspended in dimethylformamide (65 ml). To the suspension
were added potassium carbonate (100 mg, 0.72 mmol) and
1-bromo-2-fluoroethane (0.05 ml, 0.7 mmol) in order. The mixture
was stirred at room temperature for 4 hours. The solvent was
removed in vacuo, and the residue was purified by silica gel
chromatography (3% methanol/chloroform) to give the subject
compound (100 mg, yield 64%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.14 (1H, s), 7.31 (5H, m),
6.59 (br s), 4.88 (2H, s), 4.63 (1H, t, J=6.6 Hz), 4.46 (1H, t,
J=6.6 Hz), 3.31 (4H, m).
Example 56
6-Amino-9-benzyl-2-[(4-chlorobenzyl)thio]-8-hydroxypurine
##STR63##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (134 _mg, 0.49
mmol) was suspended in dimethylformamide (65 ml). To the suspension
were added potassium carbonate (100 mg, 0.72 mmol) and
4-chlorobenzyl chloride (130 mg, 0.81 mmol) in order. The mixture
was stirred at room temperature for 8 hours. The solvent was
removed in vacuo, and the residue was purified by silica gel
chromatography (5% methanol/chloroform) to give the subject
compound (74 mg, yield 38%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.13 (1H, s), 7.29 (9H, m),
6.59 (2H, br s), 4.91 (2H, s), 4.26 (s, 2H).
Example 57
6-Amino-9-benzyl-8-hydroxy-2-[(3-methoxybenzyl)thio]purine
##STR64##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (134 mg, 0.49
mmol) was suspended in dimethylformamide (60 ml). To the suspension
were added potassium carbonate (100 mg, 0.72 mmol) and
3-methoxybenzyl chloride (0.1 ml, 0.7 mmol) in order. The mixture
was stirred at room temperature for 2 hours. The solvent was
removed in vacuo, and the residue was purified by silica gel
chromatography (5% methanol/chloroform) to give the subject
compound (94 mg, yield 49%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.13 (1H, s), 7.28 (5H, m),
7.15 (1H, m), 6.96 (2H, m), 6.79 (1H, m), 6.59 (2H, br s), 4.89
(2H, s), 4.27 (2H, s), 3.68 (3H, s).
Example 58
6-Amino-9-benzyl-2-cyclohexylmethylthio-8-hydroxypurine
##STR65##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (134 mg, 0.49
mmol) was suspended in dimethylformamide (60 ml). To the suspension
were added potassium carbonate (100 mg, 0.72 mmol) and
cyclohexylmethyl bromide (0.1 ml, 0.7 mmol) in order. The mixture
was stirred at room temperature for 9 hours. The solvent was
removed in vacuo, and the residue was purified by silica gel
chromatography (3% methanol/chloroform) to give the subject
compound (93 mg, yield 51%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.09 (1H, s), 7.30 (5H, m),
6.50 (2H, br s), 4.87 (2H, s), 2.93 (2H, d, J=6.6 Hz), 1.78-0.88
(11H, m).
Example 59
6-Amino-9-benzyl-2-[(3-dimethylaminopropyl)thio]purine
##STR66##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (134 mg, 0.49
mmol) was suspended in dimethylformamide (60 ml). To the suspension
were added potassium carbonate (200 mg, 1.44 mmol) and
3-dimethylaminopropyl chloride hydrochloride (114 mg, 0.72 mmol) in
order. The mixture was stirred at room temperature for 9 hours. The
solvent was removed in vacuo, and the residue was purified by
silica gel chromatography (14% methanol/chloroform) to give the
subject compound (13 mg, yield 7%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.10 (1H, s), 7.30 (5H, m),
6.50 (2H, br s), 4.87 (2H, s), 3.00 (2H, t, J=7.6 Hz), 2.26 (2H, t,
J=7.3 Hz), 2.08 (6H, S), 1.72 (2H, m).
Example 60
3-(6-Amino-9-benzyl-8-hydroxy-2-purinyl)thio-1-propanol
##STR67##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (134 mg, 0.49
mmol) was suspended in dimethylformamide (60 ml). To the suspension
were added potassium carbonate (100 mg, 0.72 mmol) and
3-bromo-1-propanol (0.07 ml, 0.7 mmol) in order. The mixture was
stirred at room temperature for 3 hours. The solvent was removed in
vacuo, and the residue was purified by silica gel chromatography
(4% methanol/chloroform) to give the subject compound (64 mg, yield
39%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.09 (1H, s), 7.31 (5H, m),
6.51 (2H, br s), 4.87 (2H, s), 4.51 (1H, t, J=5.3 Hz), 3.48 (2H, q,
J=5.6 Hz), 3.05 (2H, t, J=6.9 Hz), 1.75 (211, m).
Example 61
6-Amino-9-benzyl-2-(3-chlorobenzyl)thio-8-hydroxypurine
##STR68##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (134 mg, 0.49
mmol) was suspended in dimethylformamide (60 ml). To the suspension
were added potassium carbonate (100 mg, 0.72 mmol) and
3-chlorobenzyl chloride (0.09 ml, 0.7 mmol) in order. The mixture
was stirred at room temperature for 5 hours. The solvent was
removed in vacuo, and the residue was purified by silica gel
chromatography (3% methanol/chloroform) to give the subject
compound (92 mg, yield 47%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.14 (1H, s), 7.45 (1H, m),
7.27 (5H, m), 6.61 (2H, br s), 4.90 (2H, s), 4.30 (2H, s).
Example 62
6-Amino-9-benzyl-8-hydroxy-2-[3-(methoxycarbonyl)propyl]-thiopurine
##STR69##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (200 mg, 0.73
mmol) was suspended in dimethylformamide (60 ml). To the suspension
were added potassium carbonate (150 mg, 1.1 mmol) and methyl
4-chlorobutylate (0.13 ml, 1.1 mmol) in order. The mixture was
stirred at room temperature for 3 hours. The solvent was removed in
vacuo, and the residue was purified by silica gel chromatography
(2% methanol/chloroform) to give the subject compound (97 mg, yield
36%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.11 (1H, s), 7.30 (5H, m),
6.53 (2H, br s), 4.88 (2H, s), 3.57 (3H, s), 3.03 (2H, t, J=7.2
Hz), 2.39 (2H, t, J=7.2 Hz), 1.86 (2H, m).
Example 63
6-Amino-9-benzyl-8-hydroxy-2-[(2-phenylethyl)thio]purine
##STR70##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (134 mg, 0.49
mmol) was suspended in dimethylformamide (60 ml). To the suspension
were added potassium carbonate (100 mg, 0.72 mmol) and
2-bromoethylbenzene (0.10 ml, 0.73 mmol) in order. The mixture was
stirred at room temperature for 4 hours. The solvent was removed in
vacuo, and the residue was purified by silica gel chromatography
(3% methanol/chloroform) to give the subject compound (117 mg,
yield 63%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.13 (1H, s), 7.28 (5H, m),
6.56 (2H, br s), 4.92 (2H, s), 3.22 (2H, m), 2.89 (2H, t, J=6.6
Hz).
Example 64
4-(6-Amino-9-benzyl-8-hydroxy-2-purinyl)thiobutyric Acid
##STR71##
To a methanol solution (5 ml) containing 500 mg of sodium hydroxide
was added
6-amino-9-benzyl-8-hydroxy-2-[(methoxycarbonylpropyl)]thiopurine
(60 mg, 0.16 mmol). The solution was refluxed under heating for 5
hours, and neutralized with 2N hydrochloric acid and then aqueous
sodium hydrogen carbonate. After removal of the solvent in vacuo,
the resulting crystals were filtered and washed with water to give
the subject compound (5 mg, yield 9%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 7.31 (5H, m), 6.90 (2H, br s),
3.02 (2H, m), 2.09 (2H, m), 1.79 (2H, m).
Example 65
6-Amino-9-benzyl-8-hydroxy-2-[(4-methoxybenzyl)thio]purine
##STR72##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (134 mg, 0.49
mmol) was suspended in dimethylformamide (60 ml). To the suspension
were added potassium carbonate (100 mg, 0.72 mmol) and
4-methoxybenzyl chloride (0.098 ml, 0.72 mmol) in order. The
mixture was stirred at room temperature for 3 hours. The solvent
was removed in vacuo, and the residue was purified by silica gel
chromatography (3% methanol/chloroform) to give the subject
compound (80 mg, yield 41%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.13 (1H, s), 7.28 (5H, m),
6.73 (2H, t, J=8.9 Hz), 6.57 (2H, br s), 4.92 (2H, s), 4.22 (2H,
s), 3.69 (3H, s).
Example 66
6-Amino-9-benzyl-2-(2-cyanoethyl)thio-8-hydroxypurine
##STR73##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (134 mg, 0.49
mmol) was suspended in dimethylformamide (60 ml). To the suspension
were added potassium carbonate (100 mg, 0.72 mmol) and
3-chloropropionitrile (65 mg, 0.73 mmol) in order. The mixture was
stirred at room temperature for 3 hours. The solvent was removed in
vacuo, and to the residue was added water. The mixture was
extracted with chloroform and the organic layer was dried on sodium
sulfate. After removal of the solvent in vacuo, the residue was
purified by silica gel chromatography (5% methanol/chloroform) to
give the subject compound (72 mg, yield 45%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.17 (1H, br s), 7.36-7.25
(5H, m), 6.63 (2H, br s), 4.90 (2H, s), 3.24 (2H, t, J=6.0 Hz),
2.88 (2H, t, J=6.0 Hz).
Example 67
6-Amino-9-benzyl-2-cyanomethylthio-8-hydroxypurine
##STR74##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (150 mg, 0.55
mmol) was suspended in dimethylformamide (10 ml). To the suspension
were added potassium carbonate (81 mg, 0.59 mmol) and
chloroacetonitrile (44 mg, 0.59 mmol) in order. The mixture was
stirred at room temperature for 3 hours. The solvent was removed in
vacuo, and to the residue was added water. The mixture was
extracted with chloroform and the organic layer was dried on sodium
sulfate. After removal of the solvent in vacuo, the residue was
purified by silica gel chromatography (1% ammonia, 5%
methanol/chloroform) to give the subject compound (58 mg, yield
25%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.21 (1H, br s), 7.41-7.24
(5H, m), 6.71 (2H, br s), 4.91 (2H, s), 4.15 (2H, s).
Example 68
6-Amino-9-benzyl-2-(3-cyanopropyl)thio-8-hydroxypurine
##STR75##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (150 mg, 0.55
mmol) was suspended in dimethylformamide (10 ml). To the suspension
were added potassium carbonate (202 mg, 1.46 mmol) and
4-chlorobutyronitrile (152 mg, 1.46 mmol) in order. The mixture was
stirred at room temperature for 3 hours. The solvent was removed in
vacuo, and to the residue was added water. The mixture was
extracted with chloroform and the organic layer was dried on sodium
sulfate. After removal of the solvent in vacuo, the residue was
purified by silica gel chromatography (5% methanol/chloroform) to
give the subject compound (71 mg, yield 29%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.13 (1H, br s), 7.36-7.22
(5H, m), 6.55 (2H, br s), 4.91 (2H, s), 3.08 (2H, t, J=7.0 Hz),
2.56 (2H, t, J=7.0 Hz), 1.91 (2H, m).
Example 69
6-Amino-9-benzyl-8-hydroxy-2-(4-methylthiomethyl)thiopurine
##STR76##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (134 mg, 0.49
mmol) was suspended in dimethylformamide (60 ml). To the suspension
were added potassium carbonate (100 mg, 0.72 mmol) and chloromethyl
methyl sulfide (0.06 ml, 0.72 mmol) in order. The mixture was
stirred at room temperature for 4 hours. The solvent was removed in
vacuo, and the residue was purified by silica gel chromatography
(3% methanol/chloroform) to give the subject compound (83 mg, yield
51%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.14 (1H, s), 7.33 (5H, m),
6.59 (2H, br s), 4.88 (2H, s), 4.30 (2H, s), 2.11 (3H, s).
Example 70
6-Amino-9-benzyl-2-(benzyloxymethyl)thio-8-hydroxypurine
##STR77##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (134 mg, 0.49
mmol) was suspended in dimethylformamide (60 ml). To the suspension
were added potassium carbonate (100 mg, 0.72 mmol) and
benzyloxymethyl chloride (0.1 ml, 0.7 mmol) in order. The mixture
was stirred at room temperature for 4 hours. The solvent was
removed in vacuo, and the residue was purified by silica gel
chromatography (3% methanol/chloroform) to give the subject
compound (90 mg, yield 47%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.15 (1H, s), 7.30 (10H, m),
6.61 (2H, br s), 4.88 (2H, s), 4.52 (2H, s).
Example 71
6-Amino-9-benzyl-8-hydroxy-2-[3-(1-piperazinyl)propyl]thiopurine
##STR78##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (134 mg, 0.49
mmol) was suspended in dimethylformamide (60 ml). To the suspension
were added potassium carbonate (100 mg, 0.72 mmol) and
1-(3-chloropropyl)piperazine (179 mg, 0.73 mmol) in order. The
mixture was stirred at room temperature for 2 hours. The solvent
was removed in vacuo, and the residue was purified by silica gel
chromatography (20% methanol/chloroform) to give the subject
compound (10 mg, yield 5%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 7.31 (5H, m), 6.62 (2H, br s),
4.87 (2H, s), 3.04-2.94 (6H, m), 2.43-2.37 (6H, m), 1.76 (2H,
m).
Example 72
6-Amino-9-benzyl-8-hydroxy-2-[2-(methylthio)ethyl]thiopurine
##STR79##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (134 mg, 0.49
mmol) was suspended in dimethylformamide (60 ml). To the suspension
were added potassium carbonate (100 mg, 0.72 mmol) and chloromethyl
ethyl sulfide (0.08 ml, 0.7 mmol) in order. The mixture was stirred
at room temperature for 2 hours. The solvent was removed in vacuo,
and the residue was purified by silica gel chromatography (3%
methanol/chloroform) to give the subject compound (28 mg, yield
16%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.11 (1H, s), 7.31 (5H, m),
6.54 (2H, br s), 4.88 (2H, s), 3.21 (2H, m), 2.73 (2H, m), 2.07
(3H, s).
Example 73
4-[(6-Amino-9-henzyl-8-hydroxy-2-purinyl)thio]butanol
##STR80##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (470 mg, 1.7
mmol) was suspended in dimethylformamide (80 ml). To the suspension
were added potassium carbonate (350 mg, 2.5 mmol) and
4-chlorobutanol (0.25 ml, 2.5 mmol) in order. The mixture was
stirred at room temperature for 4 hours. The solvent was removed in
vacuo, and the residue was purified by silica gel chromatography
(7% methanol/chloroform) to give the subject compound (29 mg, yield
5%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.09 (1H, s), 7.31 (5H, m),
6.50 (2H, br s), 4.87 (2H, s), 4.40 (1H, t, J=5.3 Hz), 3.39 (2H, q,
J=5.3 Hz), 3.02 (2H, t, J=6.9 Hz), 1.67-1.48 (4H, m).
Example 74
6-Amino-9-benzyl-8-hydroxy-2-{2-(2-methoxyethoxy)ethyl]thio}purine
##STR81##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (350 mg, 1.7
mmol) was suspended in dimethylformamide (100 ml). To the
suspension were added potassium carbonate (350 mg, 2.5 mmol) and
1-(2-chloroethoxy)-2-methoxyethane (1.04 g, 2.6 mmol) in order. The
mixture was stirred at room temperature for 2 hours. The solvent
was removed in vacuo, and the residue was purified by silica gel
chromatography (7% methanol/chloroform) to give the subject
compound (69 mg, yield 11%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.11 (1H, s), 7.30 (5H, m),
6.54 (2H, br s), 4.88 (2H, s), 3.58 (2H, t, J=6.6 Hz), 3.49 (2H, t,
J=2.6 Hz), 3.40 (2H, t, J=5.9 Hz), 3.22 (3H, s), 3.18 (2H, t, J=6.6
Hz).
Example 75
6-Amino-9-benzyl-8-hydroxy-2-{[2-(2-hydroxyethoxy)ethyl]thio}purine
##STR82##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (470 mg, 1.7
mmol) was suspended in dimethylformamide (100 ml). To the
suspension were added potassium carbonate (350 mg, 2.5 mmol) and
2-(2-chloroethoxy)ethanol (0.27 ml, 2.6 mmol) in order. The mixture
was stirred at room temperature for 4 hours. The solvent was
removed in vacuo and the residue was purified by silica gel
chromatography (7% methanol/chloroform) to give the subject
compound (159 mg, yield 27%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.11 (1H, s), 7.31 (5H, m),
6.54 (2H, br s), 4.88 (2H, s), 4.59 (1H, t, J=5.6 Hz), 3.48-3.39
(4H, m), 3.19 (2H, t, J=6.6 Hz).
Example 76
6-Amino-9-benzyl-8-hydroxy-2-{[2-(2-ethoxyethoxy)ethyl]thio}purine
##STR83##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (470 mg, 1.7
mmol) was suspended in dimethylformamide (100 ml). To the
suspension were added potassium carbonate (350 mg, 2.5 mmol) and
1-ethoxy-2-(2-bromoethoxy)ethane (505 mg, 2.6 mmol) in order. The
mixture was stirred at room temperature for 2 hours. The solvent
was removed in vacuo, and the residue was purified by silica gel
chromatography (1% methanol/chloroform) to give the subject
compound (147 mg, yield 22%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.11 (1H, s), 7.31 (5H, m),
6.54 (2H, br s), 4.88 (2H, s), 3.58 (2H, t, J=6.9 Hz), 3.51-3.36
(6H, m), 3.18 (2H, t, J=6.9 Hz), 1.07 (3H, t, J=6.9 Hz).
Example 77
6-Amino-9-benzyl-8-hydroxy-2-(3-ethoxypropyl)thiopurine
##STR84##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (80 mg, 0.65
mmol) was suspended in dimethylformamide (60 ml). To the suspension
were added potassium carbonate (150 mg, 1.1 mmol) and
2-ethoxypropyl p-toluenesulfonate (280 mg, 1.1 mmol) in order. The
mixture was stirred at room temperature for 4 hours. The solvent
was removed in vacuo, and the residue was purified by silica gel
chromatography (5% methanol/chloroform) to give the subject
compound (69 mg, yield 30%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.09 (1H, s), 7.31 (5H, m),
6.51 (2H, br s), 4.87 (2H, s), 3.44-3.34 (4H, m), 3.03 (2H, t,
J=8.9 Hz), 1.83 (2H, m), 1.08 (3H, t, J=6.9 Hz).
Example 78
6-Amino-9-benzyl-8-hydroxy-2-{[2-(2-hydroxyethylthio)ethyl]thio}purine
##STR85##
Crude 6-amino-9-benzyl-8-hydroxy-2-mercaptopurine (134 mg, 0.49
mmol) was suspended in dimethylformamide (50 ml). To the suspension
were added potassium carbonate (100 mg, 0.73 mmol) and
2-(2-chloroethyl)thioethanol (170 mg, 1.2 mmol) in order. The
mixture was stirred at room temperature for 24 hours. To the
reaction mixture were added 2N hydrochloric acid and then 28%
aqueous ammonia. The mixture was extracted with chloroform, the
organic layer was dried on magnesium sulfate and then the solvent
was removed in vacuo. The residue was purified by silica gel
chromatography (5% methanol/chloroform) to give the subject
compound (17 mg, yield 6%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.12 (1H, s), 7.31 (5H, m),
6.53 (2H, br s), 4.87 (2H, s), 4.82 (1H, t, J=5.6 Hz), 3.53 (2H, t,
J=6.6 Hz), 3.18 (2H, m), 2.77 (2H, t, J=8.2 Hz), 2.63 (2H, t, J=6.6
Hz).
Example 79
6-Amino-9-benzyl-8-hydroxy-2-(2-methoxyethyl)aminopurine
##STR86##
6-Amino-9-benzyl-8-methoxy-2-(2-methoxyethyl)aminopurine (26 mg,
0.079 mmol) in concentrated hydrochloric acid (20 ml) was stirred
at room temperature for 7 hours. The reaction mixture was made
basic with 28% aqueous ammonia. The resulting crystals were
filtered to give the subject compound (18 mg, yield 73%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 9.66 (1H, br s), 7.34-7.26
(5H, m), 6.14 (1H, t, J=4.8 Hz), 6.05 (2H, br s), 4.80 (2H, s),
3.39-3.34 (4H, m), 3.22 (3H, s).
Example 80
6-Amino-9-benzyl-2-(2-ethoxyethoxy)-8-hydroxypurine
##STR87##
6-Amino-9-benzyl-2-(2-ethoxyethoxy)-8-methoxypurine (110 mg, 0.32
mmol) in concentrated hydrochloric acid (20 ml) was stirred at room
temperature for 12 hours. The reaction mixture was evaporated in
vacuo to dryness and then 28% aqueous ammonia was added thereto.
The resulting crystals were filtered to give the subject compound
(88 mg, yield 84%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 9.98(1H, br s), 7.35-7.23(5H,
m), 6.48(2H, br s), 4.86(2H, s), 4.25(2H, t, J=4.6 Hz), 3.62(2H, t,
J=4.6 Hz), 3.45(2H, q, J=7.0 Hz), 1.11(3H, t, J=7.0 Hz).
Example 81
6-Amino-9-(4-fluorobenzyl)-8-hydroxy-2-(2-methoxyethoxy)purine
##STR88##
6-Amino-9-(4-fluorobenzyl)-8-methoxy-2-(2-methoxyethoxy)purine (49
mg, 0.14 mmol) in concentrated hydrochloric acid (20 ml) was
stirred for 12 hours. The reaction mixture was evaporated in vacuo
to dryness and then 28% aqueous ammonia was added thereto. The
resulting crystals were filtered to give the subject compound (36
mg, yield 77%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 9.97(1H, br s), 7.35(2H, m),
7.14(2H, m), 6.48(2H, br s), 4.84(2H, s), 4.27(2H, t, J=4.6 Hz),
3.59(2H, t, J=4.6 Hz), 3.28(3H, s).
Example 82
6-Amino-9-(4-fluorobenzyl)-8-hydroxy-2-methylthiopurine
##STR89##
6-amino-9-(4-fluorobenzyl)-8-hydroxy-2-mercaptopurine (200 mg,
0.687 mmol) was suspended in methanol (20 ml). To the suspension
were added potassium carbonate (190 mg, 1.37 mmol) and methyl
iodide (975 mg, 6.87 mmol) in order. The mixture was stirred at
room temperature for 30 minutes. The reaction mixture were
evaporated in vacuo to dryness and extracted with chloroform. The
organic layer was dried on sodium sulfate. After removal of the
solvent in vacuo, the residue was purified by silica gel
chromatography (3% methanol/chloroform) to give the subject
compound (63 mg, yield 30%).
.sup.1 H-NMR (DMSO-d.sub.6) .delta.: 10.11(1H, br s), 7.37(2H, m),
7.15(2H, m), 6.53(2H, br s), 4.87(2H, s), 2.43(3H, s).
Example 83
6-Amino-9-benzyl-2-(3-hydroxypropoxy)-8-hydroxypurine
##STR90##
6-Amino-9-benzyl-2-(3-hydroxypropoxy)-8-methoxypurine (83 mg, 0.25
mmol) in concentrated hydrochloric acid (5 ml) was stirred at room
temperature for 4 hours. The reaction mixture was neutralized with
28% aqueous ammonia and the resulting crystals were filtered and
washed with water to give the subject compound (40 mg, yield
51%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 9.95(1H, br s), 7.30(5H, m),
6.45(2H, br s), 4.85(2H, s), 4.50(1H, t, J=5.0 Hz), 4.20(2H, t,
J=6.0 Hz), 3.51(2H, q, J=5.0 Hz), 1.79(2H, m).
Example 84
6-Amino-9-benzyl-8-hydroxy-2-(3-ethoxypropoxy)npurine
##STR91##
6-Amino-9-benzyl-2-(3-ethoxypropoxy)-8-methoxypurine (149 mg, 0.417
mmol) in concentrated hydrochloric acid (20 ml) was stirred at room
temperature for 12 hours. The reaction mixture was evaporated in
vacuo to dryness and neutralized with 28% aqueous ammonia. The
resulting crystals were filtered to give the subject compound (112
mg, yield 78%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 9.97(1H, br s), 7.31-7.23(5H,
m), 6.45(2H, br s), 4.84(2H, s), 4.17(2H, t, J=6.6 Hz), 3.44(2H, t,
J=6.6 Hz), 3.38(2H, q, J=7.0 Hz), 1.85(2H, m), 1.08(3H, t, J=7.0
Hz).
Example 85
6-Amino-9-benzyl-8-hydroxy-2-(4-hydroxybutoxy)purine
##STR92##
6-Amino-9-benzyl-2-(4-hydroxybutoxy)-8-methoxypurine (114 mg, 0.332
mmol) in concentrated hydrochloric acid (20 ml) was stirred at room
temperature for 12 hours. The reaction mixture was evaporated in
vacuo to dryness and to the residue was added aqueous ammonia. The
resulting crystals were filtered to give the subject compound (80
mg, yield 73%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 9.95(1H, br s), 7.33-7.24(5H,
m), 6.45(2H, br s), 4.85(2H, s), 4.43(1H, t, J=5.1 Hz), 4.14(2H, t,
J=6.6 Hz), 3.42(2H, m), 1.67(2H, m), 1.50(2H, m).
Example 86
6-Amino-9-benzyl-8-hydroxy-2-(2-methylthioethoxy)purine
##STR93##
6-amino-9-benzyl-2-chloro-8-methoxypurine (190 mg, 0.56 mmol) was
added to 2-methylthioethanol (3 ml) containing sodium (110 mg, 4.78
mmol). The mixture was heated for 2 hours. Thereto were added 2N
hydrochloric acid and 28% aqueous ammonia in order. The mixture was
extracted with 3% methanol/chloroform and the organic layer was
dried on sodium sulfate. After removal of the solvent in vacuo, the
residue was purified by silica gel chromatography (3%
methanol/chloroform) to give the subject compound (59 mg, yield
27%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 9.97(1H, s), 7.30(5H, m),
6.49(2H, br s), 4.85(2H, s), 4.30(2H, t, J=7.0 Hz), 2.76(2H, t,
J=6.9 Hz), 2.10(3H, 5).
Example 87
6-Amino-9-benzyl-8-hydroxy-2-(2-hydroxyethoxy)purine
##STR94##
6-Amino-9-benzyl-2-(2-hydroxyethoxy)-8-methoxypurine (70 mg, 0.22
mmol) in concentrated hydrochloric acid (5 ml) was stirred at room
temperature for 5 hours. The reaction mixture was neutralized with
28% aqueous ammonia and the resulting crystals were filtered and
washed with water to give the subject compound (38 mg, yield
57%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 9.89(1H, br s), 7.30(5H, m),
6.46(2H, br s), 4.85(2H, s), 4.79(1H, t, J=5.6 Hz), 4.15(2H, t,
J=4.9 Hz), 3.65(2H, m).
Example 88
6-Amino-9-benzyl-8-hydroxy-2-(2-methoxyethoxy)purine
##STR95##
6-Amino-9-benzyl-8-methoxy-2-(2-methoxyethoxy)purine (21 mg, 0.064
mmol) in concentrated hydrochloric acid (20 ml) was stirred at room
temperature for 5 hours. The reaction mixture was evaporated in
vacuo to dryness and 28% aqueous ammonia was added to the residue.
The resulting crystals were filtered and washed with water to give
the subject compound (17 mg, yield 84%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 9.97(1H, br s), 7.35-7.23(5H,
m), 6.48(2H, br s), 4.86(2H, s), 4.26(2H, t, J 4.6 Hz), 3.58(2H, t,
J=4.6 Hz), 3.27(3H, s).
Example 89
6-Amino-2-(2-aminoethylthio)-9-benzyl-8-hydroxypurine
##STR96##
6-Amino-9-benzyl-8-hydroxy-2-(2-phthalimidoethylthio)purine (78 mg,
0.18 mmol) was suspended in 1M hydrazine monohydrate (10 ml). The
suspension was stirred at room temperature for 9 hours. The
reaction mixture was evaporated in vacuo to dryness and to the
residue was added 2N hydrochloric acid. Insoluble materials were
filtered off and the filtrate was neutralized with 28% aqueous
ammonia and extracted with ethyl acetate. The organic layer was
dried on magnesium sulfate and evaporated in vacuo to dryness to
give the subject compound (4 mg, yield 7%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.31(7H, m), 6.53(2H, br s),
4.88(2H, s), 3.05(2H, t, J=6.3 Hz), 2.76(2H, t, J=6.6 Hz).
Example 90
6-Amino-2-butylthio-9-(4-fluorobenzyl)-8-hydroxypurine
##STR97##
6-Amino-9-(4-fluorobenzyl)-8-hydroxy-2-thiopurine (200 mg, 0.687
mmol) and potassium carbonate (190 mg, 1.37 mmol) were dissolved in
dimethylformamide (10 ml). Butyl bromide (941 mg, 6.87 mnmol) was
added thereto and the mixture was stirred at room temperature for 4
hours. The reaction mixture was evaporated in vacuo to dryness.
Water was added to the residue and the solution was extracted with
chloroform. The organic layer was dried on magnesium sulfate and
evaporated in vacuo to dryness. The residue was purified by silica
gel chromatography (2% methanol/chloroform) to give the subject
compound (38 mg, yield 16%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 10.10(1H, br s), 7.35(2H, m),
7.14(2H, m), 6.51(2H, br s), 4.87(2H, s), 3.00(2H, t, J=7.1 Hz),
1.56(2H, m), 1.36(2H, m), 0.86(3H, t, J=7.3 Hz).
Example 91
6-Amino-9-(4-fluorobenzyl)-8-hydroxy-2-(2-methoxyethylthio)purine
##STR98##
6-Amino-9-(4-fluorobenzyl)-8-hydroxy-2-thiopurine (200 mg, 0.687
mmol) and potassium carbonate (190 mg, 1.37 mmol) were dissolved in
dimethylformamide (10 ml). 2-Methoxyethyl chloride (649 mg, 6.87
mmol) was added thereto and the mixture was stirred at room
temperature for 2 hours. The reaction mixture was evaporated in
vacuo to dryness. Water was added to the residue and the solution
was extracted with chloroform. The organic layer was dried on
magnesium sulfate and evaporated in vacuo to dryness. The residue
was purified by silica gel chromatography (2% methanol/chloroform)
to give the subject compound (50 mg, yield 21%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 10.12(1H, br s), 7.34(2H, m),
7.15(2H, m), 6.56(2H, br s), 4.87(2H, s), 3.51(2H, t, J=6.8 Hz),
3.24(3H, s), 3.20(2H, t, J=6.8 Hz).
Example 92
Inducing Activity for Biosynthesis of Interferon
Experimental Method
1) Animals
C3H/HeJ male mice (5-8 weeks) were purchased from Clea Japan
Inc.
2) Reagents
MEM (Osaka University, Microbial Research Center), FCS (GIBCO Co.
or Filtron Pty Ltd.), DMSO (Nacalai Tesque Inc.)
3) Test Compounds
About 1 mg of each test compound was accurately weighed and was
dissolved in dimethyl sulfoxide (DMSO) to prepare 1 mM or 10 mM
solution of the test compound. The solution was further diluted 500
times with a culture medium (MEM+10%FCS) and the obtained solution
was used as the sample solution in the following tests.
4) Preparation of Spleen Cells and Supernatant of the Cell Culture
Medium
After pre-feeding for a week, spleens of 2 or 3 mice were removed.
The spleen was put in PBS(-) solution and a homogenous cell
suspension was prepared by pipetting cells from the spleen. The
cell suspension was centrifuged (1200 rpm, 5 min., 4.degree. C.) to
remove the supernatant. Thereto was quickly added 0.2% ice cold
NaCl solution (4 ml) under ice cooling to make a suspension. After
30 seconds 1.6% (4 ml) ice-cold NaCl solution was added to the
suspension and the mixture was centrifuged to remove the
supernatant. The residue was suspended in PBS(-) solution (10 ml)
and the suspension was centrifuged to remove the supernatant. The
residue was suspended in 10 ml of the culture medium (MEM+10%FCS)
and the suspension was further centrifuged to remove the
supernatant.
Then, the residue was suspended in 5 ml of the culture medium to
control the viable cell number (2.times.10.sup.6 cells/ml by trypan
blue-staining). Thus obtained controlled cell suspension was poured
into a 24-well plate (0.5 ml/well) and the sample solution (0.5 ml)
was added to each well (0.5 ml/well) and the plate was incubated
(37.degree. C., 5% CO.sub.2) for 24 hours. After filtration (0.22
.mu.m) of the supernatant of the culture medium, the filtrate was
kept at -20.degree. C. as a bioassay sample.
5) Quantitative Assay of Interferon .alpha. in the Supernatant of
the Culture Medium
Immediately after L cells (Dainippon Pharm. Co.) cultured in
monolayer culture were treated with trypsin, the cells were added
to the culture medium and the cell suspension (4.times.10.sup.5
cell/ml) was prepared by pipetting. Each 100 .mu.l of the cell
suspension was poured into all wells of a 96-well plate (Sumitomo
Bakelite Co.) and the plate was incubated (37.degree. C., 5%
CO.sub.2) for about 6 hours.
Standard mouse interferon (prepared by Lee Bio Molec. Res Co.)
which was diluted by serial dilution method with a dilution plate
and the above bioassay sample were poured into an assay plate (each
of 50 .mu.l). On the other hand only the culture medium (50 .mu.l)
was added to the uninfected cell control group and to the virus
infected cell control group, respectively.
After incubation for 18 hours, the culture medium in the assay
plate was removed. A solution containing bovine vesicular
stomatitis virus [after cloning virus (3.7.times.10.sup.8 PFU/ml)
distributed by Domestic Animal Research Institute with BHK cell,
the original solution was diluted 300 times] was poured into all of
wells (100 .mu.l/well) except the virus uninfected control group.
On the other hand, only the culture medium (100 .mu.l) was added to
the virus uninfected control group.
After incubation for about 48 hours, the virus solution on the
assay plate was removed by suction. A dye solution (neutral red)
was poured into all wells (50 .mu.l/well). After incubation for 45
minutes, the dye solution was removed by suction and the wells were
washed with PBS(-) solution.
After removal of the PBS(-) solution, UV was irradiated for 10
hours to inactivate the virus. A mixture (100 .mu.l) of 0.1M
NaH.sub.2 PO.sub.4 and 99.5% ethanol (1:1) was poured into each
well and the plate was stirred by a mixer for about 5 minutes.
After that, the absorption at 540 nm was measured with a plate
reader.
6) Result of Measurement
The result was shown in Table 1. The compounds of the present
invention have inducing activity for biosynthesis of interferon.
The drug concentration in the Table means final concentration.
TABLE 1 INDUCING ACTIVITY FOR BIOSYNTHESIS OF INTERFERON Inducing
activity for biosynthesis of interferon (IU/ml) Example No. (0.1
.mu.M) (1 .mu.M) 1 93 46 16 111 26 24 3 34 25 92 50 30 80 19 42 21
13 43 18 17 44 33 14 48 28 14 54 47 18 60 31 12 66 40 23 67 36 20
79 7 18 80 25 18 81 21 18 82 29 18 88 47 26
Example 93
Activity on Cytokine Production from Mouse Lymph Node Cells
Experimental Method
1) Animals
BALB/c female mice were purchased from Japan Charlse River
(Yokohama) and female mice (8 weeks) were used.
2) Culture Medium
RPMI1640 medium "DAIGO" (Nippon Seiyaku (Tokyo)) supplemented with
10% heat-inactivated (56.degree. C., 30 min.) Fetal Bovine Serum
(characterized Code No. A-1115-L, HyClone Lab., Logan, Utah) and 50
mM 2-mercaptoethanol (Sigma, St. Louis, Mo., Code No. M-6250) were
used for the assay.
3) Test Compounds
Each test compound dissolved in DMSO (Nacalai Tesque (Kyoto) code
No. 11J) at a concentration of 100 mM was diluted to final
concentration with the medium.
4) Sensitization and Preparation of Lymph Node Cells
KLM (0.2 mg) was subcutaneously administered to mouse foot with
Freund's complete adjuvant (Difco Lab., Detroit, Mich., Code No.
3113-60-5). Ten days later popliteal lymph node was picked up and
its cell suspension was prepared.
5) Production of Cytokine by Stimulation with an Antigen
KLH (0.1 mg/ml) and the drug were added to lymph node cells
(5.times.10.sup.6 cells/ml) and the mixture was incubated at
37.degree. C. under 5% CO.sub.2 for 4 days (Corning 25850, 0.15
ml/well). Then amount of cytokine produced in the supernatant was
measured by ELISA specific to cytokine.
Amounts of interleukin 4 (IL-4) and interleukin 5 (IL-5) as a
typical Th2 type cytokine, and interferon .gamma. (IFN-.gamma.) as
a typical Th1 type cytokine were measured.
6) Method of Measurement (ELISA)
Amount of IL-4 was measured by ELISA as mentioned below. A rat
anti-mouse IL-4 antibody (Pharmingen, San Diego, Calif., Code No.
18031D, 0.5 mg/ml) as a primary antibody was diluted 250 times with
hydrogen carbonate buffer, and it was seeded to the 96-well plate
(Falcon 3912, Becton Dickinson and Company, Franklin Lakes, N.J.)
(50 ml/well) and each well was coated at 4.degree. C. overnight.
Then the plate was blocked with PBS (-) solution containing 3% BSA
(200 ml/well). After rinsing and drying the plate, the plate was
stored at -20.degree. C. until beginning to use. The supernatant of
the culture medium was added to the wells (50 ml/well) and the
plate was incubated at room temperature for 4 hours. Recombinant
mouse IL-4 (Pharmingen, Code No. 19231W) was used for preparing a
calibration curve.
After rinsing the plate, a rat anti-mouse IL-4 antibody labeled by
biotin (Pharmingen, Code No. 18042D, 0.5 mg/ml) as a secondary
antibody, which was diluted 500 times with PBS (-) solution
containing 0.1% BSA, was poured into wells (100 ml/well). The plate
was incubated at room temperature. The secondary antibody bound to
the plate was detected with streptoabidin alkaliphosphatase
(Kirkegaad & Perry Lab., Gaithersburg, Md., Code No.
15-30-00)(0.25 mg/ml, 10 ml/well). After incubation at 37.degree.
C. for 1 hour and rinsing of the plate, the coloring was done by
adding p-nitrophenyl disodium phosphate substrate (Nacalai
Tesque)(1 mg/ml, 100 ml/well). The absorption at 415 nm was
measured by a microplate reader (MTP-120 Microplatereader, Corona
Electric Co.)
Measurement of amounts of IFN-.gamma. was carried out in the same
method as mentioned above by using a rat anti-mouse IFN-.gamma.
antibody (Pharmingen, San Diego, Calif., Code No. 18181D, 0.5
mg/ml) as a primary antibody and a rat anti-mouse IFN-.gamma.
antibody labeled by biotin (Pharmingen, Code No. 18112D, 0.5 mg/ml)
as a secondary antibody. Recombinant mouse IFN-.gamma. (Pharmingen,
Code No. 19301U) was used for preparing a calibration curve.
Measurement of amounts of IL-5 was carried out in the same method
as mentioned above by using a rat anti-mouse, IL-5 antibody
(Pharmingen, San Diego, Calif., Code No. 18051D, 0.5 mg/ml) as a
primary antibody and a rat anti-mouse IL-5 antibody labeled by
biotin (Pharmingen, Code No. 18062D, 0.5 mg/ml) as a secondary
antibody. Recombinant mouse IL-5 (Pharmingen, Code No. 19241W) was
used for preparing a calibration curve. The test was carried out
three times and their average was calculated.
The test result on IL-4 was shown in the following Table 2.
TABLE 2 Activity on inhibition for production of IL-4 Example No.
Residual amount of IL-4 Inhibition (Concentration of (residual
activity Drug 10 .mu.M) (ng/nl) rate %) (%) 1 2.67 31.1 68.9 2 3.81
41.3 58.7 3 1.63 11.7 88.3 4 3.81 39.1 60.9 5 5.98 69.1 30.9 7 5.24
53.8 46.2 8 4.98 53.9 46.1 11 5.84 68.1 21.9 12 3.89 45.4 54.6 15
3.44 40.1 59.9 16 4.75 51.4 48.6 17 5.25 56.9 43.1 18 6.47 70.1
29.9 19 1.73 12.7 87.3 20 3.38 32.1 67.9 21 3.86 28.4 71.6 22 2.22
16.3 83.7 23 2.56 18.8 81.2 26 6.64 68.0 32.0 32 6.78 25.4 74.6 36
5.22 49.6 50.4 42 2.12 7.9 92.1 48 1.89 7.7 96.3 81 1.50 6.1 93.9
88 2.84 10.7 89.3 89 3.00 28.5 71.5
Example 94
Activity of the Compound on Mouse Contact Hypersensitivity Reaction
Induced by TNCB
Test Method
1) Animals
BALB/c female mice (6 weeks old) were purchased from Nippon Charles
River Co. (Kanagawa) and they were used after previously feeding
for 7 days.
2) Sensitization and Induction Method
Hair on mouse abdomen was cut and thereon was spread 7%
2,4,6-trinitrochlorobenzen (TNCB) (Tokyo Kasei (Tokyo)) in acetone
(0.1 ml/mouse) to sensitize (day 0). 6 Days later 1% TNCB in
acetone (10 ml) was spread on both sides of left auricula for
induction.
3) Administration Method
After dissolving or homogeneously suspending in acetone, a test
compound (10 ml) was spread on both sides of left auricula,
respectively. The compound was applied once one hour before
induction. As a positive control an adrenocortical hormone
(Betamethasone, Wako Chemical Co. (Osaka)) was used.
4) Method of Measurement of Thickness of Auricula
Right before and 24 hours after spreading hapten (TNCB), thickness
of left and right auriculae of each mouse was measured under
diethyl ether anesthesia by dial thickness gauge (Mitutoyo,
Tokyo).
Value of thickness of auricula was calculated by following
equation: (Value of thickness of auricula)=(thickness of spread
left auricula)-(thickness of unspread right auricula).
Inhibition rate of thickness was calculated by following
equation:
Inhibition rate of thickness={1-[(value of thickness of auricula
for 24 hours after drug application in drug-applied group)-(value
of thickness of auricula before drug application in drug-applied
group)]/[(value of thickness of auricula 24 hours after acetone
application in acetone-applied group)-(value of thickness of
auricula before drug application in acetone-applied
group)].times.100.
Result
The result was shown in Table 3.
It was observed that inhibition of thickness of auricula skin 24
hours after induction in the group of application of the drug was
superior in comparing with the group of application of acetone
substrate.
Mouse contact hypersensitivity reaction induced by TNCB is
considered as a typical model for human contact dermatitis
Therefore, the result shows that the compounds of the present
invention have therapeutic and prophylactic activity for human
contact dermatitis.
TABLE 3 Dose of Inhibition Standard Example No. drug rate (%) error
(%) 1 0.4 mg/ear 79.0 3.7 20 0.4 mg/ear 74.7 8.1 30 0.4 mg/ear 37.3
8.6 32 0.4 mg/ear 45.1 16.2 42 0.4 mg/ear 59.7 7.5 44 0.4 mg/ear
71.2 2.4 54 0.4 mg/ear 63.1 1.6 58 0.4 mg/ear 64.4 10.4 60 0.4
mg/ear 85.0 7.6 81 0.4 mg/ear 79.0 5.2 88 0.4 mg/ear 39.1 10.4 90
0.4 mg/ear 80.3 3.2 Betamethasone 0.001 mg/ear 91.0 2.2 Acetone
substrate 0.0 9.8
Example 95
Activity of the Compound on Mouse Contact Hypersensitivity Reaction
Induced by TNCB
Test Method
The test on compound of Example 79 was carried out in the same
method as in Example 94.
Result
The result was shown in Table 4.
It was observed that inhibition of thickness of auricula skin 24
hours after induction in the group of application of compound of
Example 79 was superior in comparing with the group of application
of acetone substrate.
Mouse contact hypersensitivity reaction induced by the TNCB is
considered as a typical model for human contact dermatitis
Therefore, the result shows that compound of Example 79 has
therapeutic and prophylactic activity for human contact
dermatitis.
TABLE 4 Dose of Inhibition Standard Example No. drug rate (%) error
(%) 79 0.4 mg/ear 68.9 2.3 Betamethasone 0.001 mg/ear 64.4 5.3
Acetone substrate 0.0 4.5
Example 96
Activity Against Ear Edema Reaction Induced by Arachidonic Acid
Test Method
1) Animal
1) Animals
BALB/c female mice (6 weeks) was purchased from Nippon Charles
River Co. (Kanagawa) and they were used after previously feeding
until 7 weeks old.
2) Administration Method
After weighing compound of Example 44 it was suspended in acetone
(Kanto Kagaku Co.) (20 mg/ml). The suspension (10 .mu.l) was spread
on both sides of left auricula of mouse under anesthesia with
diethyl ether, respectively. As a control acetone (10 .mu.l) was
spread on both sides of left auricula of another mouse,
respectively.
2) Spread of Arachidonic Acid
Two hours after spread of compound of Example 44 or acetone, 10%
arachidonic acid (CAYMAN CHEMICAL, Michigan) (10 .mu.l) was spread
on both sides of left auricula, respectively.
3) Measurement of Interdermal Reaction
One hour after spreading 10% arachidonic acid, thickness of both
left and right auriculae was measured under anesthesia with diethyl
ether by Dial thickness gauge (Mitutoyo, Tokyo).
Value of thickness of the auricula was calculated by following
equation:
Inhibition rate of the thickness was calculated by following
equation:
Inhibition rate of thickness={1-[(value of thickness of auricula 1
hour after drug application in drug-applied group)-(value of
thickness of auricula before drug application in drug-applied
group)]/[(value of thickness of auricula 1 hour after acetone
application in acetone-applied group)-(value of thickness of
auricula 1 hour before acetone application in acetone-applied
group)].times.100.
Result
The result was shown in Table 5.
It was observed that inhibition of thickness of auricular skin 1
hour after induction in the group of application of compound of
Example 44 was superior in comparing with the group of application
of acetone substrate.
The result indicates that the compounds of the present invention
inhibit dermal inflammatory induced by arachidonic acid.
It is suggested that inflammatory mediators, that is, arachidonic
acid metabolites, such as prostaglandins, leucotrienes and
hydroxyeicosatetraenoic acids participate in dermal inflammatory
disease such as psoriasis, UV dermatitis, mastocytosis and dermal
cancer.
It was suggested that the compounds of the present invention are
useful as therapeutic agents for diseases related to arachidonic
acid metabolites.
TABLE 5 Dose of Inhibition Standard Example No. drug rate (%) error
(%) 44 0.4 mg/ear 94.03 5.97 Acetone substrate 0.0 25.37
Reference Example 1
6-Amino-2-chloropurine
##STR99##
A solution of 2,6-dichloropurine 0.5 g (2.7 mmol) in 30%
ammonia-methanol solution was heated at 100.degree. C. in autoclave
for 12 hours. The solution was condensed to give the subject
compound. The compound may be used in next reaction without further
purification.
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 8.13(1H, s), 7.66(2H, br
s).
Reference Example 2
6-Amino-9-benzyl-2-chloropurine
##STR100##
6-Amino-2-chloropurine (295 mg) and potassium carbonate (0.55 g,
4.0 mmol) were suspended in DMF (10 ml). Benzyl bromide (0.17 ml,
1.4 mmol) was added thereto and the mixture was stirred at room
temperature for 4 hours. After condensing the suspension in vacuo,
to the residue was added brine and the mixture was extracted with
chloroform. The organic layer was washed the mixture was with
brine, dried on magnesium sulfate, filtered and the solvent in the
filtrate was evaporated in vacuo. The residue was purified with
silica gel chromatography (5% methanol/chloroform) and
recrystallized from ethanol to give the subject compound (200 mg,
yield 58%). m.p. 216-218.degree. C.
UV.lambda..sub.max (EtOH): 265.7 nm; .sup.1 H-NMR(DMSO-d.sub.6)
.delta.: 8.26(1H, s), 7.81(2H, br s), 7.31(5H, m), 5.34(2H, s).
Reference Example 3
6-Amino-9-benzyl-2-methylthiopurine
##STR101##
6-Amino-9-benzyl-2-chloropurine (100 mg, 0.39 mmol) and sodium
methylthiolate (270 mg, 3.9 mmol) were mixed in DMF (10 ml) and
then the mixture was stirred at 10.degree. C. for 3.5 hours. To the
reaction mixture was added brine and the mixture was extracted with
ethyl acetate. The organic layer was dried on magnesium sulfate,
filtered and the solvent in the filtrate was evaporated in vacuo.
The residue was purified with silica gel chromatography (1%
methanol/chloroform) to give the subject compound (64 mg, yield
61%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.63(1H, s), 7.34(5H, m),
5.45(2H,br s), 5.31(2H, s), 2.58(3H, s).
Reference Example 4
6-Amino-9-benzyl-2-ethylthiopurine
##STR102##
To DMF suspension (10 ml) containing sodium hydride (300 mg, 7.5
mmol, 60% in mineral oil) were added ethanethiol (2 ml, 27 mmol)
and 6-amino-9-benzyl-2-chloropurine (100 mg, 0.39 mmol) in order.
The mixture was stirred under heating at 110.degree. C. for 3.5
hours. Brine was added thereto and the mixture was extracted with
ethyl acetate. The organic layer was dried on magnesium sulfate,
filtered and the solvent in the filtrate was evaporated. The
residue was purified with silica gel chromatography (1%
methanol/chloroform) to give the subject compound (90 mg, yield
82%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.64(1H, s), 7.33(5H, m),
5.91(2H, br s), 5.29(2H, s), 3.17(2H, q, J=7.3 Hz), 1.39(3H, t,
J=7.3 Hz).
Reference Example 5
6-Amino-9-benzyl-2-propylthiopurine
##STR103##
To DMF suspension (50 ml) containing sodium hydride (917 mg, 23
mmol, 60% in mineral oil) were added propanethiol (5.0 ml, 55 mmol)
and 6-amino-9-benzyl-2-chloropurine (500 mg, 1.9 mmol) in order.
The mixture was stirred under heating at 110.degree. C. for 2.5
hours. Brine was added thereto and the mixture was extracted with
ethyl acetate. The organic layer was dried on magnesium sulfate,
filtered and the solvent in the filtrate was evaporated. The
residue was purified with silica gel chromatography (1%
methanol/chloroform) to give the subject compound (505 mg, yield
87%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.64(1H, s), 7.32(5H, m),
6.09(2H, br s), 5.28(2H, s), 3.14(2H, t, J=7.3 Hz), 1.76(2H, m),
1.03(3H, t, J=7.3 Hz).
Reference Example 6
6-Amino-9-benzyl-2-(isopropylthio)purine
##STR104##
To DMF suspension (10 ml) containing sodium hydride (300 mg, 7.5
mmol 60% in mineral oil) were added 2-propanethiol (1.0 ml, 11
mmol) and 6-amino-9-benzyl-2-chloropurine (160 mg, 0.62 mmol) in
order. The mixture was stirred under heating at 100.degree. C. for
2.5 hours. Brine was added thereto and the mixture was extracted
with chloroform. The organic layer was dried on magnesium sulfate,
filtered and the solvent in the filtrate was evaporated in vacuo.
The residue was purified with silica gel chromatography (1%
methanol/chloroform) to give the subject compound (112 mg, yield
61%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.64(1H, s), 7.32(5H, m),
5.49(2H, br s), 5.29(2H, s), 3.98(1H, m), 1.43(6H, d, J=6.6
Hz).
Reference Example 7
6-Amino-9-benzyl-2-butylthiopurine
##STR105##
6-Amino-9-benzyl-2-chloropurine (310 mg, 1.2 mmol) and sodium
butylthiolate (670 mg, 6.0 mmol) were mixed in DMF (30 ml) and then
the mixture was stirred under heating at 100.degree. C. for 4.5
hours. To the reaction mixture was added brine and the mixture was
extracted with ethyl acetate. The organic layer was dried on
magnesium sulfate, filtered and the solvent in the filtrate was
evaporated in vacuo. The residue was purified with silica gel
chromatography (0.5% methanol/chloroform) to give the subject
compound (194 mg, yield 52%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.63(1H, s), 7.35(5H, m),
5.54(2H, br s), 5.29(2H, s), 3.17(2H, t, J=7.3 Hz), 1.72(2H, m),
1.48(2H, m), 0.93(3H, t, J=7.6 Hz).
Reference Example 8
6-Amino-9-benzyl-2-(isobutylthio)purine
##STR106##
To DMF suspension (10 ml) containing sodium hydride (300 mg, 7.5
mmol, 60% in mineral oil) were added 2-methylpropane-1-thiol (1 ml,
11 mmol) and 6-amino-9-benzyl-2-chloropurine (200 mg, 0.77 mmol) in
order. The mixture was stirred under heating at 100.degree. C. for
5 hours. Brine was added thereto and the mixture was extracted with
chloroform. The organic layer was dried on magnesium sulfate,
filtered and the solvent in the filtrate was evaporated in vacuo.
The residue was purified with silica gel chromatography (1%
methanol/chloroform) to give the subject compound (76 mg, yield
31%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.63(1H, s), 7.32(5H, m),
5.46(2H, br s), 5.29(2H, s), 3.08(d, 2H, J=6.9 Hz), 2.00(1H, m),
1.04(6H, d, J=6.6 Hz).
Reference Example 9
6-Amino-9-benzyl-2-(sec-butylthio)purine
##STR107##
To DMF suspension (10 ml) containing sodium hydrate (300 mg, 7.5
mmol, 60%in mineral oil) were added 2-butanethiol (1 ml, 11 mmol)
and 6-amino-9-benzyl-2-chloropurine (200 mg, 0.77 mmol) in order.
The mixture was stirred under heating at 100.degree. C. for 5
hours. Brine was added thereto and the mixture was extracted with
chloroform. The organic layer was dried on magnesium sulfate,
filtered and the solvent in the filtrate was evaporated in vacuo.
The residue was purified with silica gel chromatography (1%
methanol/chloroform) to give the subject compound (85 mg, yield
35%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.63(1H, s), 7.32(5H, m),
5.46(2H, br s), 5.29(2H, s), 3.85(1H, m), 1.75(2H, m), 1.42(3H, d,
J=6.9 Hz), 1.03(3H, t, J=7.6 Hz).
Reference Example 10
6-Amino-9-benzyl-2-pentylthiopurine
##STR108##
To DMF suspension (10 ml) containing sodium hydride (277 mg, 6.9
mmol, 60% in mineral oil) were added 1-pentanethiol (2 ml, 16 mmol)
and 6-amino-9-benzyl-2-chloropurine (100 mg, 0.39 mmol) in order.
The mixture was stirred under heating at 110.degree. C. for 4
hours. Brine was added thereto and the mixture was extracted with
ethyl acetate. The organic layer was dried on magnesium sulfate,
filtered and the solvent in the filtrate was evaporated in vacuo.
The residue was purified with silica gel chromatography (1%
methanol/chloroform) to give the subject compound (102 mg, yield
81%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.64(1H, s), 7.33(5H, m),
5.77(2H, br s), 5.29(2H, s), 3.16(2H, t, J=7.3 Hz), 1.75(2H, m),
1.33-1.46(4H, m), 0.89(3H, t, J=7.3 Hz).
Reference Example 11
6-Amino-9-benzyl-2-(3-methylbutyl)thiopurine
##STR109##
To DMF suspension (10 ml) containing sodium hydride (300 mg, 7.5
mmol, 60% in mineral oil) were added 3-methylbutane-1-thiol (1 ml,
8.0 mmol) and 6-amino-9-benzyl-2-chloropurine (200 mg, 0.77 mmol)
in order. The mixture was stirred under heating at 100.degree. C.
for 2.5 hours. Brine was added thereto and the mixture was
extracted with chloroform. The organic layer was dried on magnesium
sulfate, filtered and the solvent in the filtrate was evaporated in
vacuo. The residue was purified with silica gel chromatography (1%
methanol/chloroform) to give the subject compound (120 mg, yield
48%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.63(1H, s), 7.32(5H, m),
5.44(2H, br s), 5.29(2H, s), 3.17(2H, t, J=7.9 Hz), 1.64(3H, m),
0.94(6H, d, J=6.6 Hz).
Reference Example 12
6-Amino-9-benzyl-2-(2-methylbutyl)thiopurine
##STR110##
To DMF suspension (10 ml) containing sodium hydride (300 mg, 7.5
mmol, 60% in mineral oil) were added 2-methylbutane-1-thiol (1 ml,
8.0 mmol) and 6-amino-9-benzyl-2-chloropurine (200 mg, 0.77 mmol)
in order. The mixture was stirred under heating at 100.degree. C.
for 4.5 hours. Brine was added thereto and the mixture was
extracted with chloroform. The organic layer was dried on magnesium
sulfate, filtered and the solvent in the filtrate was evaporated in
vacuo. The residue was purified with silica gel chromatography (1%
methanol/chloroform) to give the subject compound (80 mg, yield
32%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.63(1H, s), 7.32(5H, m),
5.50(2H, br s), 5.30(2H, s), 3.26(1H, q, J=5.9 Hz), 2.99(1H, q,
J=7.6 Hz), 1.78(1H, m), 1.55(1H, m), 1.28(1H, m), 1.02(3H, d,
J=11.9 Hz), 0.92(3H, t, J=11.8 Hz).
Reference Example 13
6-Amino-9-benzyl-2-cyclohexylthiopurine
##STR111##
To DMF suspension (10 ml) containing sodium hydride (256 mg, 6.4
mmol, 60% in mineral oil) were added cyclohexanethiol (2 ml, 16
mmol) and 6-amino-9-benzyl-2-chloropurine (100 mg, 0.39 mmol) in
order. The mixture was heated at 100.degree. C. for 3.5 hours.
Brine was added thereto and the mixture was extracted with ethyl
acetate. The organic layer was dried on magnesium sulfate,
filtrated and the solvent in the filtrate was evaporated in vacuo.
The residue was purified with silica gel chromatography (1%
methanol/chloroform) to give the subject compound (112 mg, yield
86%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.65(1H, s), 7.33(5H, m),
5.86(2H, br s), 5.28(2H, m), 3.75-3.87(1H, m), 2.11-2.17(2H, m),
1.25-1.67(8H, m).
Reference Example 14
6-Amino-9-benzyl-2-phenylthiopurine
##STR112##
6-Amino-9-benzyl-2-chloropurine (200 mg, 0.77 mmol) and sodium
thiophenolate (2 g, 15 mmol) were mixed in DMF (12 ml) and then the
mixture was stirred under heating at 100.degree. C. for 7.5 hours.
To the reaction mixture was added brine and the mixture was
extracted with ethyl acetate. The organic layer was dried on
magnesium sulfate, filtered and the solvent in the filtrate was
evaporated in vacuo. The residue was purified with silica gel
chromatography (0.5% methanol/chloroform) to give the subject
compound (228 mg, yield 89%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.65-7.70(3H, m), 7.41-7.45(3H,
m), 7.28-7.33(3H, m), 7.15-7.20(2H, m), 5.54(2H, br s), 5.09(2H,
s).
Reference Example 15
6-Amino-9-benzyl-2-(p-tolylthio)purine
##STR113##
To DMF suspension (10 ml) containing sodium hydride (300 mg, 7.5
mmol, 60% in mineral oil) were added p-toluenethiol (1.9 g, 15
mmol) and 6-amino-9-benzyl-2-chloropurine (100 mg, 0.39 mmol) in
order. The mixture was stirred under heating at 100.degree. C. for
3 hours. Brine was added thereto and the mixture was extracted with
chloroform. The organic layer was dried on magnesium sulfate,
filtrated and the solvent in the filtrate was evaporated in vacuo.
The residue was purified with silica gel chromatography (1%
methanol/chloroform) to give the subject compound (124 mg, yield
93%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.62(1H, s), 7.55(2H, d, J=8.2
Hz), 7.15-7.31(7H, m), 5.61(2H, br s), 5.10(2H, s), 2.40(3H,
s).
Reference Example 16
6-Amino-9-benzyl-2-(2-naphthylthio)purine
##STR114##
To DMF suspension (20 ml) containing sodium hydride (800 mg, 20
mmol, 60% in mineral oil) were added 2-naphthalenethiol (3.8 g, 24
mmol) and 6-amino-9-benzyl-2-chloropurine (200 mg, 0.77 mmol) in
order. The mixture was stirred under heating at 100.degree. C. for
10.5 hours. Brine was added thereto and the mixture was extracted
with chloroform. The organic layer was dried on magnesium sulfate,
filtrated and the solvent in the filtrate was evaporated in vacuo.
The residue was purified with silica gel chromatography (0.5%
methanol/chloroform) to give the subject compound (244 mg, yield
83%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 8.17(1H, s), 7.52-7.92(7H, m),
7.06-7.30(5H, m), 5.63(2H, br s), 5.04(2H, s).
Reference Example 17
6-Amino-9-benzyl-2-benzylthiopurine
##STR115##
To DMF suspension (10 ml) containing sodium hydride (410 mg, 10
mmol, 60% in mineral oil) were added .alpha.-toluenethiol (1.7 ml,
14 mmol) and 6-amino-9-benzyl-2-chloropurine (100 mg, 0.39 mmol) in
order. The mixture was stirred under heating at 100.degree. C. for
4.5 hours. Brine was added thereto and the mixture was extracted
with ethyl acetate. The organic layer was dried on magnesium
sulfate, filtrated and the solvent in the filtrate was evaporated
in vacuo. The residue was purified with silica gel chromatography
(0.5% methanol/chloroform) to give the subject compound (97 mg,
yield 73%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.64(1H, s), 7.22-7.45(10H, m),
5.48(2H, br s), 5.31(2H, s), 4.43(2H, s).
Reference Example 18
6-Amino-9-benzyl-8-bromo-2-methylthiopurine
##STR116##
6-Amino-9-benzyl-2-methylthiopurine (100 mg, 0.37 mmol) and bromine
(0.5 ml) were dissolved in 100 ml of methylene chloride and the
solution was stirred at room temperature for 3 hours. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated, dried on magnesium sulfate and filtered. The
solvent in the filtrate was evaporated in vacuo. The residue was
purified with silica gel chromatography (0.5% methanol/chloroform)
to give the subject compound (10 mg, yield 8%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.34(5H, m), 5.64(2H, br s),
5.33(2H, s), 2.57(3H, s).
Reference Example 19
6-Amino-9-benzyl-8-bromo-2-ethylthiopurine
##STR117##
6-Amino-9-benzyl-2-ethylthiopurine (214 mg, 0.75 mmol) and bromine
(0.5 ml) were dissolved in 100 ml of methylene chloride and the
solution was stirred at room temperature for 7 hours. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated, dried on magnesium sulfate and filtered. The
solvent in the filtrate was evaporated in vacuo. The residue was
purified with silica gel chromatography (1% methanol/chloroform) to
give the subject compound (43 mg, yield 16%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.32(5H, m), 5.82(2H, br s),
5.32(2H, s), 3.16(2H, q, J=7.3 Hz), 1.39(3H, t, J=7.3 Hz).
Reference Example 20
6-Amino-9-benzyl-8-bromo-2-propylthiopurine
##STR118##
6-Amino-9-benzyl-2-propylthiopurine (290 mg, 0.97 mmol) and bromine
(0.7 ml) were dissolved in 160 ml of methylene chloride and the
solution was stirred at room temperature for 4.5 hours. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated, dried on magnesium sulfate and filtered. The
solvent in the filtrate was evaporated in vacuo. The residue was
purified with silica gel chromatography (1% methanol/chloroform) to
give the subject compound (58 mg, yield 16%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.35(5H, m), 5.70(2H, br s),
5.32(2H, s), 3.13(2H, t, J=7.6 Hz), 1.76(2H, m), 1.04(3H, t, J=7.6
Hz).
Reference Example 21
6-Amino-9-benzyl-8-bromo-2-(isopropylthio)purine
##STR119##
6-Amino-9-benzyl-2-(isopropylthio)purine (60 mg, 0.20 mmol) and
bromine (0.4 ml) were dissolved in 85 ml of methylene chloride and
the solution was stirred at room temperature for 2 hours. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated, dried on magnesium sulfate and filtered. The
solvent in the filtrate was evaporated in vacuo. The residue was
purified with silica gel chromatography (1% methanol/chloroform) to
give the subject compound (20 mg, yield 26%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.34(5H, m), 5.72(2H, br s),
5.32(2H, s), 3.96(1H, m), 1.42(6H, d, J=7.0 Hz).
Reference Example 22
6-Amino-9-benzyl-8-bromo-2-butylthiopurine
##STR120##
6-Amino-9-benzyl-2-butylthiopurine (163 mg, 0.52 mmol) and bromine
(0.6 ml) were dissolved in 180 ml of methylene chloride and the
solution was stirred at room temperature for 4.5 hours. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated, dried on magnesium sulfate and filtered. The
solvent in the filtrate was evaporated in vacuo. The residue was
purified with silica gel chromatography (1% methanol/chloroform) to
give the subject compound (35 mg, yield 17%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.34(5H, m), 5.81(2H, br s),
5.32(2H, s), 3.15(2H, t, J=7.3 Hz), 1.72(2H, m), 1.45(2H, m),
0.92(3H, t, J=7.6 Hz).
Reference Example 23
6-Amino-9-benzyl-8-bromo-2-(isobutylthio)purine
##STR121##
6-Amino-9-benzyl-2-(isobutylthio)purine (60 mg, 0.19 mmol) and
bromine (0.4 ml) were dissolved in 85 ml of methylene chloride and
the solution was stirred at room temperature for 2 hours. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated, dried on magnesium sulfate and filtered. The
solvent in the filtrate was evaporated in vacuo. The residue was
purified with silica gel chromatography (chloroform) to give the
subject compound (20 mg, yield 27%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.34(5H, m), 5.59(2H, br s),
5.32(2H, s), 3.07(2H, t, J=6.6 Hz), 1.96(1H, m), 1.04(6H, d, J=6.6
Hz).
Reference Example 24
6-Amino-9-benzyl-8-bromo-2-(sec-butylthio)purine
##STR122##
6-Amino-9-benzyl-2-(sec-butylthio)purine (60 mg, 0.19 inmol) and
bromine (0.4 ml) were dissolved in 85 ml of methylene chloride and
the solution was stirred at room temperature for 2 hours. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated, dried on magnesium sulfate and filtered. The
solvent in the filtrate was evaporated in vacuo. The residue was
purified with silica gel chromatography (chloroform) to give the
subject compound (53 mg, yield 71%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.34(5H, m), 5.45(2H, br s),
5.32(2H, s), 3.83(1H, m), 1.63(2H, m), 1.42(3H, d, J=7.0 Hz),
1.03(3H, t, J=7.3 Hz).
Reference Example 25
6-Amino-9-benzyl-8-bromo-2-pentylthiopurine
##STR123##
6-Amino-9-benzyl-2-pentylthiopurine (260 mg, 0.79 mmol) and bromine
(0.5 ml) were dissolved in 100 ml of methylene chloride and the
solution was stirred at room temperature for 7 hours. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated, dried on magnesium sulfate and filtered. The
solvent of the filtrate was evaporated in vacuo. The residue was
purified with silica gel chromatography (1% methanol/chloroform) to
give the subject compound (49 mg, yield 15%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.33(5H, m), 5.95(2H, br s),
5.31(2H, s), 3.14(2H, t, J=7.3 Hz), 1.74(2H, m), 1.27-1.47(4H, m),
0.88(3H, t, J=7.3 Hz).
Reference Example 26
6-Amino-9-benzyl-8-bromo-2-(3-methylbutyl)thiopurine
##STR124##
6-Amino-9-benzyl-2-(3-methylbutyl)thiopurine (260 mg, 0.79 mmol)
and bromine (0.5 ml) were dissolved in 100 ml of methylene chloride
and the solution was stirred at room temperature for 7 hours.
Aqueous sodium thiosulfate was added to the reaction mixture. The
organic layer was separated, dried on magnesium sulfate and
filtered. The solvent in the filtrate was evaporated in vacuo. The
residue was purified with silica gel chromatography (1%
methanol/chloroform) to give the subject compound (49 mg, yield
15%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.33(5H, m), 5.52(2H, br s),
5.30(2H, s), 3.15(2H, t, J=7.9 Hz), 1.61-1.76(3H, m), 0.92(6H, t,
J=6.2 Hz).
Reference Example 27
6-Amino-9-benzyl-8-bromo-2-(2-methylbutyl)thiopurine
##STR125##
6-Amino-9-benzyl-2-(2-methylbutyl)thiopurine (60 mg, 0.18 mmol) and
bromine (0.4 ml) were dissolved in 90 ml of methylene chloride and
the solution was stirred at room temperature for 7 hours. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated, dried on magnesium sulfate and filtered. The
solvent in the filtrate was evaporated in vacuo. The residue was
purified with silica gel chromatography (1% methanol/chloroform) to
give the subject compound (39 mg, yield 53%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.33(5H, m), 5.44(2H, br s),
5.32(2H, s), 3.24(1H, q, J=7.9 Hz), 2.98(1H, q, J=7.3 Hz), 1.75(1H,
m), 1.52(1H, m), 1.28(1H, m), 1.01(3H, d, J=6.6 Hz), 0.91(3H, t,
J=7.3 Hz).
Reference Example 28
6-Amino-9-benzyl-8-bromo-2-cyclohaxylthiopurine
##STR126##
6-Amino-9-benzyl-2-cyclohexylthiopurine (178 mg, 0.52 mmol) and
bromine (0.4 ml) were dissolved in 90 ml of methylene chloride and
the solution was stirred at room temperature for 7 hours. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated, dried on magnesium sulfate and filtered. The
solvent in the filtrate was evaporated in vacuo. The residue was
purified with silica gel chromatography (0.5% methanol/chloroform)
to give the subject compound (86 mg, yield 40%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.30-7.45(5H, m), 5.69(2H, br s),
5.31(2H, s), 3.80(1H, m), 2.10(2H, m), 1.25-1.78(8H, m).
Reference Example 29
6-Amino-9-benzyl-8-bromo-2-phenylthiopurine
##STR127##
6-Amino-9-benzyl-2-phenylthiopurine (95 mg, 0.28 mmol) and bromine
(0.4 ml) were dissolved in 150 ml of methylene chloride and the
solution was stirred at room temperature for 4.5 hours. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated, dried on magnesium sulfate and filtered. The
solvent in the filtered was evaporated in vacuo. The residue was
purified with silica gel chromatography (0.5% methanol/chloroform)
to give the subject compound (25 mg, yield 22%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.65-7.68(2H, m), 7.42-7.44(3H,
m), 7.20-7.28(5H, m), 5.49(2H, br s), 5.09(2H, s).
Reference Example 30
6-Amino-9-benzyl-8-bromo-2-(p-tolylthio)purine
##STR128##
6-Amino-9-benzyl-2-(p-tolylthio)purine (86 mg, 0.37 mmol) and
bromine (0.4 ml) were dissolved in 120 ml of methylene chloride and
the solution was stirred at room temperature for 4 hours. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated, dried on magnesium sulfate and filtered. The
solvent in the filtrate was evaporated in vacuo. The residue was
purified with silica gel chromatography (0.5% methanol/chloroform)
to give the subject compound (20 mg, yield 19%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.55(2H, d, J=7.9 Hz),
7.20-7.28(7H, m), 5.40(2H, br s), 5.10(2H, s), 2.41(3H, s).
Reference Example 31
6-Amino-9-benzyl-8-bromo-2-(2-naphthylthio)purine
##STR129##
6-Amino-9-benzyl-2-(2-naphthylthio)purine (221 mg, 0.58 mmol) and
bromine (0.4 ml) were dissolved in 160 ml of methylene chloride and
the solution was stirred at room temperature for 5.5 hours. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated, dried on magnesium sulfate and filtered. The
solvent in the filtrate was evaporated in vacuo. The residue was
purified with silica gel chromatography (0.5% methanol/chloroform)
to give the subject compound (118 mg, yield 44%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 8.42(1H, d, J=8.3 Hz),
7.80-7.87(3H, m), 7.52-7.66(2H, m), 7.04-7.21(6H, m), 5.56(2H, br
s), 5.00(2H, s).
Reference Example 32
6-Amino-9-benzyl-2-benzylthio-8-bromopurine
##STR130##
6-Amino-9-benzyl-2-benzylthiopurine (176 mg, 0.51 mmol) and bromine
(1 ml) were dissolved in 160 ml of methylene chloride and the
solution was stirred at room temperature for 4 hours. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated, dried on magnesium sulfate and filtered. The
solvent in the filtrate was evaporated in vacuo. The residue was
purified with silica gel chromatography (0.5% methanol/chloroform)
to give the subject compound (19 mg, yield 9%).
.sup.1 H-NMR(CDCl.sub.3) .delta.: 7.21-7.39(10H, m), 5.50(2H, br
s), 5.33(2H, s), 4.41(2H, m).
Reference Example 33
6-Amino-9-benzyl-2-methoxypurine
##STR131##
6-Amino-9-benzyl-2-chloropurine (200 mg, 0.77 mmol) and sodium
methylate (208 mg, 3.85 mmol) were dissolved in methanol (20 ml)
and then the solution was refluxed on heating under stirring for 30
hours. The reaction mixture was evaporated in vacuo to dryness. To
the residue was added water and the mixture was extracted with
chloroform. The organic layer was dried on sodium sulfate and
evaporated in vacuo to dryness. The residue was purified with
silica gel chromatography (2% methanol/chloroform) to give the
subject compound (151 mg, yield 77%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 8.05(1H, s), 7.37-7.25(7H, m),
5.26(2H, s), 3.81(3H, s).
Reference Example 34
6-Amino-9-benzyl-2-ethoxypurine
##STR132##
6-Amino-9-benzyl-2-chloropurine (200 mg, 0.77 mmol) and sodium
ethylate (262 mg, 3.85 mmol) were dissolved in ethanol (20 ml) and
then the solution was refluxed on heating under stirring for 20
hours. The reaction mixture was evaporated in vacuo to dryness. To
the residue was added water and the mixture was extracted with
chloroform. The organic layer was dried on sodium sulfate and
evaporated in vacuo to dryness. The residue was purified with
silica gel chromatography (2% methanol/chloroform) to give the
subject compound (151 mg, yield 73%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 8.04(1H, s), 7.37-7.21(7H, m),
5.25(2H, s), 4.25(2H, q, J=7.1 Hz), 1.27(3H, t, J=7.1 Hz).
Reference Example 35
6-Amino-9-benzyl-2-propoxypurine
##STR133##
6-Amino-9-benzyl-2-chloropurine (200 mg, 0.77 mmol) and sodium
propylate (316 mg, 3.85 mmol) were dissolved in 1-propanol (20 ml)
and then the solution was refluxed on heating under stirring for 3
hours. The reaction mixture was evaporated in vacuo to dryness. To
the residue was added water and the mixture was extracted with
chloroform. The organic layer was dried on sodium sulfate and
evaporated in vacuo to dryness. The residue was purified with
silica gel chromatography (2% methanol/chloroform) to give the
subject compound (162 mg, yield 74%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 8.04(1H, s), 7.37-7.21(7H, m),
5.26(2H, s), 4.16(2H, t, J=6.6 Hz), 1.68(2H, m), 0.95(3H, t, J=7.3
Hz).
Reference Example 36
6-Amino-9-benzyl-2-butoxypurine
##STR134##
6-Amino-9-benzyl-2-chloropurine (200 mg, 0.77 mmol) and sodium
butylate (370 mg, 3.85 mmol) were dissolved in 1-butanol (20 ml)
and then the solution was refluxed on heating under stirring for 2
hours. The reaction mixture was evaporated in vacuo to dryness. To
the residue was added water and the mixture was extracted with
chloroform. The organic layer was dried on sodium sulfate and
evaporated in vacuo to dryness. The residue was purified with
silica gel chromatography (2% methanol/chloroform) to of give the
subject compound (131 mg, yield 54%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 8.03(1H, s), 7.37-7.21(7H, m),
5.25(2H, s), 4.20(2H, t, J=6.4 Hz), 1.65(2H, m), 1.39(2H, m),
0.92(3H, t, J=7.3 Hz).
Reference Example 37
6-Amino-9-benzyl-2-pentoxypurine
##STR135##
6-Amino-9-benzyl-2-chloropurine (150 mg, 0.58 mmol) and sodium
pentylate (318 mg, 2.89 mmol) were dissolved in 1-pentanol (50 ml)
and then the solution was stirred under heating at 130.degree. C.
for 5 hours. The reaction mixture was evaporated in vacuo to
dryness. To the residue was added water and the mixture was
extracted with chloroform. The organic layer was dried on sodium
sulfate and evaporated in vacuo to dryness. The residue was
purified with silica gel chromatography (2% methanol/chloroform) to
give the subject compound (103 mg, yield 57%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 8.03(1H, s), 7.37-7.25(5H, m),
7.20(2H, br s), 5.26(2H, s), 4.20(2H, t, J=6.6 Hz), 1.67(2H, m),
1.33(4H, m), 0.88(3H, t, J=6.6 Hz).
Reference Example 38
6-Amino-9-benzyl-8-bromo-2-methoxypurine
##STR136##
6-Amino-9-benzyl-2-methoxypurine (118 mg, 0.46 mmol) and bromine
(0.5 ml) were dissolved in 50 ml of methylene chloride and the
solution was stirred at room temperature for 5 hours. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated, dried on sodium sulfate and evaporated in
vacuo to dryness. The residue was purified with silica gel
chromatography (1% methanol/chloroform) to give the subject
compound (90 mg, yield 58%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.48 (2H, br s), 7.39-7.24 (5H,
m), 5.26 (2H, s), 3.82 (3H, s).
Reference Example 39
6-Amino-9-benzyl-8-bromo-2-ethoxypurine
##STR137##
6-Amino-9-benzyl-2-ethoxypurine (143 mg, 0.53 mmol) and bromine
(0.5 ml) were dissolved in 50 ml of methylene chloride and the
solution was stirred at room temperature for 5 hours. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated and dried on sodium sulfate, filtered and
evaporated in vacuo to dryness. The residue was purified with
silica gel chromatography (1% methanol/chloroform) to give the
subject compound (42 mg, yield 23%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.43(2H, br s), 7.38-7.24(5H,
m), 5.25(2H, s), 4.26(2H, q, J=7.1 Hz), 1.28(3H, t, J=7.1 Hz).
Reference Example 40
6-Amino-9-benzyl-8-bromo-2-propoxypurine
##STR138##
6-Amino-9-benzyl-2-ethoxypurine (134 mg, 0.473 mmol) and bromine
(0.5 ml) were dissolved in 50 ml of methylene chloride and the
solution was stirred at room temperature for 5 hours. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated and dried on sodium sulfate and evaporated in
vacuo to dryness. The residue was purified with silica gel
chromatography (1% methanol/chloroform) to give the subject
compound (55 mg, yield 32%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.43(2H, br s), 7.38-7.23(5H,
m), 5.25(2H, s), 4.16(2H, t, J=6.6 Hz), 1.70(2H, m), 0.94(3H, t,
J=7.3 Hz).
Reference Example 41
6-Amino-9-benzyl-8-bromo-2-butoxypurine
##STR139##
6-Amino-9-benzyl-2-butoxypurine (120 mg, 0.404 mmol) and bromine
(0.5 ml) were dissolved in 50 ml of methylene chloride and the
solution was stirred at room temperature for 5 hours. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated and dried on sodium sulfate and evaporated in
vacuo to dryness. The residue was purified with silica gel
chromatography (1% methanol/chloroform) to give the subject
compound (97 mg, yield 64%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.44(2H, br s), 7.37-7.23(5H,
m), 5.26(2H, s), 4.21(2H, t, J=6.4 Hz), 1.64(2H, m), 1.39(2H, m),
0.91(3H, t, J=7.3 Hz).
Reference Example 42
6-Amino-9-benzyl-8-bromo-2-pentoxypurine
##STR140##
6-Amino-9-benzyl-2-pentoxypurine (95 mg, 0.305 mmol) and bromine
(0.5 ml) were dissolved in 100 ml of methylene chloride and the
solution was stirred at room temperature for 1 hour. Aqueous sodium
thiosulfate was added to the reaction mixture. The organic layer
was separated and dried on sodium sulfate and evaporated in vacuo
to dryness. The residue was purified with silica gel chromatography
(1% methanol/chloroform) to give the subject compound (78 mg, yield
82%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.42(2H, br s), 7.37-7.22(5H,
m), 5.25(2H, s), 4.19(2H, t, J=6.4 Hz), 1.66(2H, m), 1.33(4H, m),
0.88(3H, t, J=6.8 Hz).
Reference Example 43
2,6-Diamino-9-benzylpurine
##STR141##
2,6-Diaminopurine (5.00 g, 33.3 mmol) and potassium carbonate (6.91
g, 50.0 mmol) were suspended in DMF (250 ml). Benzyl bromide (8.55
g, 50 mmol) was added thereto and the mixture was stirred at room
temperature for 5 hours. After condensing the reaction mixture in
vacuo, to the residue was added water and the mixture was extracted
with chloroform. The organic layer was dried on sodium sulfate,
filtered and the solvent in the filtrate was evaporated in vacuo.
The residue was purified with silica gel chromatography (5%
methanol/chloroform) to give the subject compound (1.56 g, yield
19%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.78 (1H, s), 7.36-7.21 (5H,
m), 6.69 (2H, br s), 5.80 (2H, br s), 5.19 (2H, s).
Reference Example 44
6-Amino-9-benzyl-2-methylaminopurine
##STR142##
6-Amino-9-benzyl-2-chloropurine (200 mg, 0.77 mmol) and 40%
methylamine/methanol solution (50 ml) were heated at 120.degree. C.
for 20 hours in autoclave. The reaction mixture was condensed in
vacuo. To the residue was added 5N aqueous sodium hydroxide and the
solution was extracted with chloroform. The organic layer was dried
on sodium sulfate, filtered and the solvent in the filtrate was
evaporated in vacuo. The residue was purified with silica gel
chromatography (2% methanol/chloroform) to give the subject
compound (163 mg, yield 83%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.78 (1H, s), 7.36-7.26 (5H,
m), 6.68 (2H, br s), 6.20 (1H, q, J=4.8 Hz), 5.19 (2H, s), 2.76
(3H, d, J=4.8 Hz).
Reference Example 45
6-Amino-9-benzyl-2-ethylaminopurine
##STR143##
6-Amino-9-benzyl-2-chloropurine (200 mg, 0.77 mmol) and aqueous
ethylamine (50 ml) were heated at 120.degree. C. for 20 hours in
autoclave. The reaction mixture was condensed in vacuo. To the
residue was added 5N aqueous sodium hydroxide and the mixture was
extracted with chloroform. The organic layer was dried on sodium
sulfate, filtered and the solvent in the filtrate was evaporated in
vacuo. The residue was purified with silica gel chromatography (2%
methanol/chloroform) to give the subject compound (147 mg, yield
71%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.78 (1H, s), 7.36-7.26 (5H,
m), 6.65 (2H, br s), 6.22 (1H, t, J=5.7 Hz), 5.18 (2H, s), 3.26
(2H, m), 1.09 (3H, t, J=7.1 Hz).
Reference Example 46
6-Amino-9-benzyl-2-propylaminopurine
##STR144##
6-Amino-9-benzyl-2-chloropurine (10 mg, 0.385 mmol) and propylamine
(228 mg, 3.85 mmol) in methanol (50 ml) were heated at 120.degree.
C. for 10 hours in autoclave. The reaction mixture was condensed in
vacuo and to the residue was added 5N aqueous sodium hydroxide,
followed by extraction with chloroform. The organic layer was dried
on sodium sulfate, filtered and the solvent in the filtrate was
evaporated in vacuo. The residue was purified with silica gel
chromatography (2% methanol/chloroform) to give the subject
compound (99 mg, yield 91%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.78 (1H, s), 7.33-7.26 (5H,
m), 6.64 (2H, br s), 6.25 (1H, t, J=5.7 Hz), 5.17 (2H, s), 3.18
(2H, m), 1.50 (2H, m), 0.87 (3H, t, J=7.5 Hz).
Reference Example 47
6-Amino-9-benzyl-2-butylamtnopurine
##STR145##
6-Amino-9-benzyl-2-chloropurine (100 mg, 0.38S5 mmol) and
butylamine (282 mg, 3.85 mmol) in methanol (50 ml) were heated at
120.degree. C. for 10 hours in autoclave. The reaction mixture was
condensed in vacuo. To the residue was added 5N aqueous sodium
hydroxide and the solution was extracted with chloroform. The
organic layer was dried on sodium sulfate, filtered and the solvent
in the filtrate was evaporated in vacuo. The residue was purified
with silica gel chromatography (2% methanol/chloroform) to give the
subject compound (113 mg, yield 99%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.82 (1H, s), 7.34-7.26 (5H,
m), 6.81 (2H, br s), 6.34 (1H, t, J=6.2 Hz), 5.18 (2H, s), 3.24
(2H, m), 1.49 (2H, m), 1.31 (2H, m), 0.88 (3H, t, J=7.3 Hz).
Reference Example 48
6-Amino-9-benzyl-2-pentylaminopurine
##STR146##
6-Amino-9-benzyl-2-chloropurine (100 mg, 0.385 mmol) and
pentylamine (336 mg, 3.85 mmol) suspended in 1-butanol (10 ml) were
heated at 100.degree. C. for 10 hours in autoclave. The reaction
mixture was condensed in vacuo. To the residue was added 1N aqueous
sodium hydroxide and the solution was extracted with chloroform.
The organic layer was dried on sodium sulfate, filtered and the
solvent in the filtrate was evaporated in vacuo. The residue was
purified with silica gel chromatography (2% methanol/chloroform) to
give the subject compound (83 mg, yield 70%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.79 (1H, s), 7.32-7.26 (5H,
m), 6.62 (2H, br s), 6.21 (1H, t, J=6.0 Hz), 5.17 (2H, s),
3.25-3.18 (2H,m), 1.52-1.47 (2H, m), 1.30-1.26 (4H, m), 0.86 (3H,
t, J=6.6 Hz).
Reference Example 49
6-Amino-9-benzyl-2-(isopropylamino)puring
##STR147##
6-Amino-9-benzyl-2-chloropurine (100 mg, 0.385 mmol) and
isopropylamine (228 mg, 3.85 mmol) suspended in 1-butanol (10 ml)
were heated at 100.degree. C. for 10 hours in autoclave. The
reaction mixture was condensed in vacuo. To the residue was added
1N aqueous sodium hydroxide and the solution was extracted with
chloroform. The organic layer was dried on sodium sulfate, filtered
and the solvent in the filtrate was evaporated in vacuo. The
residue was purified with silica gel chromatography (2%
methanol/chloroform) to give the subject compound (89 mg, yield
82%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.79 (1H, s), 7.36-7.26 (5H,
m), 6.62 (2H, br s), 6.00 (1H, d, J=8.9 Hz), 5.17 (2H, s),
4.10-3.98 (1H, m), 1.11 (6H, d, J=6.6 Hz).
Reference Example 50
6-Amino-9-benzyl-2-(isobutylamino)purine
##STR148##
6-Amino-9-benzyl-2-chloropurine (100 mg, 0.385 mmol) and
isobutylamine (288 mg, 3.85 mmol) suspended in 1-butanol (10 ml)
were heated at 100.degree. C. for 10 hours in autoclave. The
reaction mixture was condensed in vacuo. To the residue was added
1N aqueous sodium hydroxide and the solution was extracted with
chloroform. The organic layer was dried on sodium sulfate, filtered
and the solvent in the filtrate was evaporated in vacuo. The
residue was purified with silica gel chromatography (2%
methanol/chloroform) to give the subject compound (89 mg, yield
78%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.79 (1H, s), 7.33-7.26 (5H,
m), 6.62 (2H, br s), 6.28 (1H, t, J=6.0 Hz), 5.17 (2H, s), 3.07
(2H, dd, J=6.0, 6.0 Hz), 1.89-1.79 (1H, m), 0.87 (6H, d, J=6.8
Hz).
Reference Example 51
6-Amino-9-benzyl-2-(sec-butylamino)purine
##STR149##
6-Amino-9-benzyl-2-chloropurine (100 mg, 0.385 mmol) and
sec-butylamine (282 mg, 3.85 mmol) suspended in 1-butanol (10 ml)
were heated at 100.degree. C. for 10 hours in autoclave. The
reaction mixture was condensed in vacuo. To the residue was added
1N aqueous sodium hydroxide and the mixture was extracted with
chloroform. The organic layer was dried on sodium sulfate, filtered
and the solvent in the filtrate was evaporated in vacuo. The
residue was purified with silica gel chromatography (2%
methanol/chloroform) to give the subject compound (71 mg, yield
62.8%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.78 (1H, s), 7.33-7.26 (5H,
m), 6.60 (2H, br s), 5.97 (1H, d, J=8.4 Hz), 5.17 (2H, s),
3.90-3.85 (1H, m), 1.54-1.38 (2H, m), 1.08 (3H, d, J=6.4 Hz), 0.85
(3H, t, J=7.3 Hz).
Reference Example 52
6-Amino-9-benzyl-2-(2,2-dimethylpropyl)aminopurine
##STR150##
6-Amino-9-benzyl-2-chloropurine (100 mg, 0.385 mmol) and
neo-pentylamine (336 mg, 3.85 mmol) suspended in 1-butanol (10 ml)
were heated at 100.degree. C. for 10 hours in autoclave. The
reaction mixture was condensed in vacuo. To the residue was added
1N aqueous sodium hydroxide and the solution was extracted with
chloroform. The organic layer was dried on sodium sulfate, filtered
and the solvent in the filtrate was evaporated in vacuo. The
residue was purified with silica gel chromatography (2%
methanol/chloroform) to give the subject compound (88 mg, yield
74%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.78 (1H, s), 7.32-7.24 (5H,
m), 6.61 (2H, br s), 6.08 (1H, t, J=6.2 Hz), 5.17 (2H, s), 3.15
(2H, d, J=6.2 Hz), 0.87 (9H, s).
Reference Example 53
6-Amino-9-benzyl-2-benzylaminopurine
##STR151##
6-Amino-9-benzyl-2-chloropurine (200 mg, 0.77 mmol) and benzylamine
(825 mg, 7.70 mmol) in 1-butanol (10 ml) were refluxed on heating
for 8 hours. The reaction mixture was condensed in vacuo. To the
residue was added 5N aqueous sodium hydroxide and the solution was
extracted with chloroform. The organic layer was dried on sodium
sulfate, filtered and the solvent in the filtrate was evaporated in
vacuo. The residue was purified with silica gel chromatography (2%
methanol/chloroform) to give the subject compound (171 mg, yield
67%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.80 (1H, s), 7.34-7.15 (10H,
m), 6.86 (1H, t, J=6.4 Hz), 6.69 (2H, br s), 5.15 (2H, s), 4.47
(2H, d, J=6.4 Hz).
Reference Example 54
6-Amino-9-benzyl-2-cyclohexylaminopurine
##STR152##
6-Amino-9-benzyl-2-chloropurine (200 mg, 0.77 mmol) and
cyclohexylamine (764 mg, 7.70 mmol) in 1-butanol (10 ml) were
refluxed on heating for 60 hours. The reaction mixture was
condensed in vacuo. To the residue was added 5N aqueous sodium
hydroxide and the solution was extracted with chloroform. The
organic layer was dried on sodium sulfate, filtered and the solvent
in the filtrate was evaporated in vacuo. The residue was purified
with silica gel chromatography (2% methanol/chloroform) to give the
subject compound (115 mg, yield 46%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.79 (1H, s), 7.33-7.26 (5H,
m), 6.60 (2H, br s), 6.00 (1H, d, J=8.1 Hz), 5.16 (2H, s), 3.71
(1H, m), 1.86 (2H, m), 1.72 (2H, m), 1.68 (1H, m), 1.31-1.14 (5H,
m).
Reference Example 55
6-Amino-2-anilino-9-benzylpurine
##STR153##
6-Amino-9-benzyl-2-chloropurine (100 mg, 0.385 mmol) and aniline
(359 mg, 3.85 mmol) in 1-butanol (10 ml) were refluxed on heating
for 20 hours. The reaction mixture was condensed in vacuo. To the
residue was added 5N aqueous sodium hydroxide and the mixture was
extracted with chloroform. The organic layer was dried on sodium
sulfate, filtered and the solvent in the filtrate was evaporated in
vacuo. The residue was purified with silica gel chromatography (2%
methanol/chloroform) to give the subject compound (108 mg, yield
89%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 8.88 (1H, s), 7.98 (1H, s),
7.81 (2H, d, J=7.9 Hz), 7.38-7.25 (5H, m), 7.20 (2H, t, J=8.3 Hz),
6.95 (2H, br s), 6.83 (1H, t, J=7.3 Hz), 5.29 (2H, s).
Reference Example 56
6-Amino-9-benzyl-2-dimethylaminopurine
##STR154##
6-Amino-9-benzyl-2-chloropurine (100 mg, 0.385 mmol) and aqueous
dimethylamine (30 ml) were heated at 120.degree. C. for 15 hours in
autoclave. The reaction mixture was condensed in vacuo. To the
residue was added 5N aqueous sodium hydroxide and the solution was
extracted with chloroform. The organic layer was dried on sodium
sulfate, filtered and the solvent in the filtrate was evaporated in
vacuo. The residue was purified with silica gel chromatography (2%
methanol/chloroform) to give the subject compound (90 mg, yield
87%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.82 (1H, s), 7.37-7.25 (5H,
m), 6.73 (2H, br s), 5.19 (2H, s), 3.07 (6H, s).
Reference Example 57
6-Amino-9-benzyl-(N-benzylmethylamino)purine
##STR155##
6-Amino-9-benzyl-2-chloropurine (100 mg, 0.385 mmol) and
N-methylbenzylamine (467 mg, 3.85 mmol) in 1-butanol (30 ml) were
refluxed on heating for 10 hours. The reaction mixture was
condensed in vacuo. To the residue was added 5N aqueous sodium
hydroxide and the mixture was extracted with chloroform. The
organic layer was dried on sodium sulfate, filtered and the solvent
in the filtrate was evaporated in vacuo. The residue was purified
with silica gel chromatography (2% methanol/chloroform) to give the
subject compound (97 mg, yield 73%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.85 (1H, s), 7.35-7.19 (10H,
m), 6.78 (2H, br s), 5.18 (2H, s), 4.85 (2H, s), 3.05 (3H, s).
Reference Example 58
2,6-Diamino-9-benzyl-8-bromopurine
##STR156##
2,6-Diamino-9-benzylpurine (1.00 g, 4.16 mmol) and bromine (1 ml)
were dissolved in 100 ml of methylene chloride and the solution was
stirred at room temperature for 5 hours. Aqueous sodium thiosulfate
was added to the reaction mixture. The organic layer was separated,
dried on sodium sulfate and filtered. The solvent in the filtrate
was evaporated in vacuo. The residue was purified with silica gel
chromatography (1% methanol/chloroform) to give the subject
compound (0.62 g, yield 47%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.37-7.16 (5H, m), 6.92 (2H, br
s), 5.99 (2H, br s), 5.18 (2H, s).
Reference Example 59
6-Amino-9-benzyl-8-bromo-2-methylaminopurine
##STR157##
6-Amino-9-benzyl-2-methylaminopurine (75 mg, 0.30 mmol) and bromine
(0.5 ml) were dissolved in 50 ml of methylene chloride and the
solution was stirred at room temperature for 1 hour. Aqueous sodium
thiosulfate was added to the reaction mixture. The organic layer
was separated, dried on sodium sulfate and filtered. The solvent of
the filtrate was evaporated in vacuo. The residue was purified with
silica gel chromatography (1% methanol/chloroform) to give the
subject compound (73 mg, yield 74%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.38-7.22 (5H, m), 6.90 (2H, br
s), 6.39 (1H, q, J=4.8 Hz), 5.18 (2H, s), 2.75 (3H, d, J=4.8
Hz).
Reference Example 60
6-Amino-9-benzyl-8-bromo-2-ethylaminopurine
##STR158##
6-Amino-9-benzyl-2-ethylaminopurine (75 mg, 0.28 mmol) and bromine
(0.5 ml) were dissolved in 50 ml of methylene chloride and the
solution was stirred at room temperature for 1 hour. Aqueous sodium
thiosulfate was added to the reaction mixture. The organic layer
was separated, dried on sodium sulfate and filtered. The solvent in
the filtrate was evaporated in vacuo. The residue was purified with
silica gel chromatography (1% methanol/chloroform) to give the
subject compound (63 mg, yield 65%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.37-7.23 (5H, m), 6.87 (2H, br
s), 6.41 (1H, t, J=5.5 Hz), 5.17 (2H, s), 3.25 (2H, m), 1.08 (3H,
t, J=7.1 Hz).
Reference Example 61
6-Amino-9-benzyl-8-bromo-2-propylaminopurine
##STR159##
6-Amino-9-benzyl-2-propylaminopurine (87 mg, 0.31 mmol) and bromine
(0.5 ml) were dissolved in 50 ml of methylene chloride and the
solution was stirred at room temperature for 1 hour. Aqueous sodium
thiosulfate was added to the reaction mixture. The organic layer
was separated, dried on sodium sulfate and filtered. The solvent in
the filtrate was evaporated in vacuo. The residue was purified with
silica gel chromatography (1% methanol/chloroform) to give the
subject compound (95 mg, yield 85%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.37-7.23 (5H, m), 6.85 (2H, br
s), 6.44 (1H, t, J=5.7 Hz), 5.17 (2H, s), 3.18 (2H, m), 1.50 (2H,
m), 0.86 (3H, t, J=7.3 Hz).
Reference Example 62
6-Amino-9-benzyl-8-bromo-2-butylaminopurine
##STR160##
6-Amino-9-benzyl-2-butylaminopurine (101 mg, 0.34 mmol) and bromine
(0.5 ml) were dissolved in 50 ml of methylene chloride and the
solution was stirred at room temperature for 1 hour. Aqueous sodium
thiosulfate was added to the reaction mixture. The organic layer
was separated, dried on sodium sulfate and filtered. The solvent in
the filtrate was evaporated in vacuo. The residue was purified with
silica gel chromatography (1% methanol/chloroform) to give the
subject compound (116 mg, yield 91%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.36-7.26 (5H, m), 6.85 (2H, br
s), 6.42 (1H, t, J=6.2 Hz), 5.17 (2H, s), 3.22 (2H, m), 1.46 (2H,
m), 1.30 (2H, m), 0.87 (3H, t, J=7.3 Hz).
Reference Example 63
6-Amino-9-benzyl-8-bromo-2-pentylaminopurine
##STR161##
6-Amino-9-benzyl-2-pentylaminopurine (70 mg, 0.23 mmol) and bromine
(0.5 ml) were dissolved in 50 ml of methylene chloride and the
solution was stirred at room temperature for 1 hour. Aqueous sodium
thiosulfate was added to the reaction mixture. The organic layer
was separated, dried on sodium sulfate and filtered. The solvent in
the filtrate was evaporated in vacuo. The residue was purified with
silica gel chromatography (1% methanol/chloroform) to give the
subject compound (80 mg, yield 91%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.37-7.24 (5H, m), 6.84 (2H, br
s), 6.41 (1H, t, J=5.7 Hz), 5.17 (2H, s), 3.25-3.18 (2H,m),
1.52-1.47 (2H, m), 1.29-1.24 (4H, m), 0.85 (3H, t, J=6.9 Hz).
Reference Example 64
6-Amino-9-benzyl-8-bromo-2-(isopropylamino)purine
##STR162##
6-Amino-9-benzyl-2-(isopropylamino)purine (71 mg, 0.25 mmol) and
bromine (0.5 ml) were dissolved in 50 ml of methylene chloride and
the solution was stirred at room temperature for 1 hour. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated, dried on sodium sulfate and filtered. The
solvent in the filtrate was evaporated in vacuo. The residue was
purified with silica gel chromatography (1% methanol/chloroform) to
give the subject compound (73 mg, yield 81%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.38-7.23 (5H, m), 6.84 (2H, br
s), 6.21 (1H, d, J=8.1 Hz), 5.17 (2H, s), 4.09-3.99 (1H, m), 1.11
(6H, d, J=6.4 Hz).
Reference Example 65
6-Amino-9-benzyl-8-bromo-2-(isobutylamino)purine
##STR163##
6-Amino-9-benzyl-2-(isobutylamino)purine (75 mg, 0.25 mmol) and
bromine (0.5 ml) were dissolved in 50 ml of methylene chloride and
the solution was stirred at room temperature for 1 hour. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated, dried on sodium sulfate and filtered. The
solvent of the filtrate was evaporated in vacuo. The residue was
purified with silica gel chromatography (1% methanol/chloroform) to
give the subject compound (62 mg, yield 65%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.37-7.27 (5H, m), 6.83 (2H, br
s), 6.47 (1H, t, J=6.0 Hz), 5.17 (2H, s), 3.06 (2H, dd, J=6.0, 6.0
Hz), 1.88-1.78 (1H, m), 0.86 (6H, d, J=6.8 Hz).
Reference Example 66
6-Amino-9-benzyl-8-bromo-2-(sec-butylamino)purine
##STR164##
6-Amino-9-benzyl-2-(sec-butylamino)purine (58 mg, 0.20 mmol) and
bromine (0.5 ml) were dissolved in 50 ml of methylene chloride and
the solution was stirred at room temperature for 1 hour. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated, dried on sodium sulfate and filtered. The
solvent in the filtrate was evaporated in vacuo. The residue was
purified with silica gel chromatography (1% methanol/chloroform) to
give the subject compound (57 mg, yield 78%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.35-7.25 (5H, m), 6.80 (2H, br
s), 6.16 (1H, d, J=8.6 Hz), 5.15 (2H, s), 3.88-3.81 (1H, m),
1.50-1.36 (2H, m), 1.05 (3H, d, J=6.4 Hz), 0.83 (3H, t, J=7.3
Hz).
Reference Example 67
6-Amino-9-benzyl-8-bromo-2-(2,2-dimethylpropyl)aminopurine
##STR165##
6-Amino-9-benzyl-2-(2,2-dimethyipropyl)aminopurine (69 mg, 0.22
mmol) and bromine (0.5 ml) were dissolved in 50 ml of methylene
chloride and the solution was stirred at room temperature for 1
hour. Aqueous sodium thiosulfate was added to the reaction mixture.
The organic layer was separated, dried on sodium sulfate and
filtered. The solvent in the filtrate was evaporated in vacuo. The
residue was purified with silica gel chromatography (1%
methanol/chloroform) to give the subject compound (75 mg, yield
87%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.36-7.23 (5H, m), 6.82 (2H, br
s), 6.29 (1H, t, J=6.2 Hz), 5.18 (2H, s), 3.14 (2H, d, J=6.2 Hz),
0.86 (9H, s).
Reference Example 68
6-Amino-9-benzyl-2-(N-benzylamino)-8-bromopurine
##STR166##
6-Amino-9-benzyl-2-(N-benzylamino)purine (60 mg, 0.18 mmol) and
bromine (0.5 ml) were dissolved in 50 ml of methylene chloride and
the solution was stirred at room temperature for 1 hour. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated, dried on sodium sulfate and filtered. The
solvent in the filtrate was evaporated in vacuo. The residue was
purified with silica gel chromatography (1% methanol/chloroform) to
give the subject compound (37 mg, yield 50%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.33-7.20 (10H, m), 7.05 (1H,
t, J=6.4 Hz), 6.91 (2H, br s), 5.15 (2H, s), 4.46 (2H, d, J=6.4
Hz).
Reference Example 69
6-Amino-9-benzyl-8-bromo-2-cyclohexylaminopurine
##STR167##
6-Amino-9-benzyl-2-cyclohexylaminopurine (100 mg, 0.31 mmol) and
bromine (0.5 ml) were dissolved in 50 ml of methylene chloride and
the solution was stirred at room temperature for 1 hour. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated, dried on sodium sulfate and filtered. The
solvent in the filtrate was evaporated in vacuo. The residue was
purified with silica gel chromatography (1% methanol/chloroform) to
give the subject compound (105 mg, yield 84%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.37-7.27 (5H, m), 6.81 (2H, br
s), 6.20 (1H, d, J=7.9 Hz), 5.16 (2H, s), 3.68 (1H, m), 1.87 (2H,
m), 1.69 (2H, m), 1.58 (1H, m), 1.30-1.12 (5H, m).
Reference Example 70
6-Amino-2-anilino-9-benzyl-8-bromopurine
##STR168##
6-Amino-2-anilino-9-benzylpurine (87 mg, 0.31 mmol) was dissolved
in a mixture of methylene chloride (50 ml) and acetic acid (10 ml).
To the solution were added sodium acetate (105 mg, 1.28 mmol) and
bromine (0.5 ml), and the mixture was stirred at room temperature
for 3 hour. Aqueous sodium thiosulfate was added to the reaction
mixture. The organic layer was separated, washed with aqueous
saturated sodium hydrogen carbonate, dried on sodium sulfate and
filtered. The solvent in the filtrate was evaporated in vacuo. The
residue was purified with silica gel chromatography (1%
methanol/chloroform) to give the subject compound (93 mg, yield
92%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 8.30 (1H, d, J=8.8 Hz), 7.82
(1H, d, J=2.4 Hz), 7.71 (1H, s), 7.49 (1H, dd, J=9.0, 2.4 Hz),
7.39-7.25 (9H, m), 5.27 (2H, s).
Reference Example 71
6-Amino-9-benzyl-8-bromo-2-dimethylaminopurine
##STR169##
6-Amino-9-benzyl-2-dimethylaminopurine (66 mg, 0.25 mmol) was
dissolved in a mixture of methylene chloride (50 ml) and acetic
acid (10 ml). To the solution were added sodium acetate (202 mg,
2.46 mmol) and bromine (0.5 ml), and the mixture was stirred at
room temperature for 1 hour. Aqueous sodium thiosulfate was added
to the reaction mixture. The organic layer was separated, washed
with aqueous saturated sodium hydrogen carbonate, dried on sodium
sulfate and filtered. The solvent in the filtrate was evaporated in
vacuo. The residue was purified with silica gel chromatography (1%
methanol/chloroform) to give the subject compound (68 mg, yield
80%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.38-7.25 (5H, m), 6.95 (2H, br
s), 5.19 (2H, s), 3.07 (6H, s).
Reference Example 72
6-Amino-9-benzyl-2-(N-benzylmethylamino)-8-bromopurine
##STR170##
6-Amino-9-benzyl-2-(N-benzylmethylamino)purine (77 mg, 0.22 mmol)
and bromine (0.5 ml) were dissolved in 50 ml of methylene chloride
and the solution was stirred at room temperature for 1 hour.
Aqueous sodium thiosulfate was added to the reaction mixture. The
organic layer was separated, dried on sodium sulfate and filtered.
The solvent in the filtrate was evaporated in vacuo. The residue
was purified with silica gel chromatography (1%
methanol/chloroform) to give the subject compound (91 mg, yield
96%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.31-7.19 (10H, m), 7.00 (2H,
br s), 5.18 (2H, s), 4.84 (2H, s), 3.05 (3H, s).
Reference Example 73
5-Amino-1-benzyl-4-cyano-2-hydroxyimidazole
##STR171##
Benzylisocyanate (25 g, 188 mmol) and N,N-diisopropylethylamine
(23.5 ml, 130 mmol) were added to aminomalononitrile
p-toluenesulfonate (45 g, 178 mmol) suspended in tetrahydrofuran.
The mixture was stirred at room temperature for 14 hours and then
the solvent was removed in vacuo. To the residue was added ethyl
acetate and the solution was washed and the organic layer was dried
on magnesium sulfate. The solvent was removed in vacuo. To the
residue was added tetrahydrofuran and 1N aqueous sodium hydroxide.
The solution was stirred at 50.degree. C. for 20 minutes, and
neutralized with 15% aqueous potassium hydrogen sulfide. The
resulting crystals were filtered and dried to give the subject
compound (41 g, 106%). The crude product was used for next reaction
without further purification.
.sup.1 H NMR (.delta., DMSO-d.sub.6): 9.91 (s, 1H), 7.31 (m, 5H),
6.51(br s, 2H), 4.76 (s, 2H).
Reference Example 74
1-Amino-9-benzyl-8-hydroxy-2-mercaptpurine
##STR172##
Crude 5-amino-1-benzyl-4-cyano-2-hydroxyimidazole (31.3 g, 146
mmol) of Reference Example 73 was suspended in tetrahydrofuran and
to the suspension was dropped benzoylisothiocyanate (41 ml, 305
mmol). After stirring overnight, the solvent was removed in vacuo
and to the residue was added ether. The crystals were filtered and
dissolved in a mixture of tetrahydrofuran and 2N aqueous sodium
hydroxide. The solution was refluxed for 50 hours and then,
neutralized with 10% aqueous potassium hydrogen sulfide. The
resulting crystals were filtered to give a mixture (27.8 g) of the
subject compound and 6-amino-7-benzyl-8-hydroxy-2-mercaptpurine.
The mixture was recrystallized from ethyl acetate to give only the
subject compound.
.sup.1 H NMR (.delta., DMSO-d.sub.6): 12.10 (br s, 1H), 10.06 (br
s, 1H), 7.36-7.24 (m, 5H), 6.74 (br s, 2H), 4.85 (s, 2H).
Reference Example 75
6-Amino-9-benzyl-2,8-dimethoxypurine
##STR173##
To 6-amino-9-benzyl-8-bromo-2-methoxypurine (125 mg, 0.374 mmol) in
10 ml of methanol was added 10N aqueous sodium hydroxide (50 ml)
and the solution was refluxed under heating for 2 hours. The
reaction mixture was concentrated in vacuo to dryness and to the
residue was added water. The mixture was extracted with chloroform
and the organic layer was dried on sodium sulfate. After removal of
the solvent the residue was purified with silica gel chromatography
(1% methanol/chloroform) to give the subject compound (83 mg, yield
78%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.73-7.23(5H, m), 6.90(2H, br
s), 5.05(2H, s), 4.04(3H, s), 3.78(3H, s).
Reference Example 76
6-Amino-9-benzyl-2-ethoxy-8-methoxypurine
##STR174##
To 6-amino-9-benzyl-8-bromo-2-ethoxypurine (35 mg, 0.101 mmol) in 5
ml of methanol was added 10N aqueous sodium hydroxide (50 ml) and
the solution was refluxed under heating for 2 hours. The reaction
mixture was concentrated in vacuo to dryness and to the residue was
added water. The mixture was extracted with chloroform and the
organic layer was dried on sodium sulfate. After removal of the
solvent the residue was purified with silica gel chromatography (1%
methanol/chloroform) to give the subject compound (22 mg, yield
73%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.73-7.23(5H, m), 6.86(2H, br
s), 5.04(2H, s), 4.22(2H, q, J=7.1 Hz), 4.04(3H, s), 1.27(3H, t,
J=7.1 Hz).
Reference Example 77
6-Amino-9-benzyl-8-methoxy-2-propoxypurine
##STR175##
To 6-amino-9-benzyl-8-bromo-2-propoxypurine (123 mg, 0.339 mmol) in
10 ml of methanol was added 10N aqueous sodium hydroxide (50 ml)
and the solution was refluxed under heating for 2 hours. The
reaction mixture was concentrated in vacuo to dryness and to the
residue was added water. The mixture was extracted with chloroform
and the organic layer was dried on sodium sulfate. After removal of
the solvent the residue was purified with silica gel chromatography
(1% methanol/chloroform) to give the subject compound (99 mg, yield
93%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.36-7.22(5H, m), 6.86(2H, br
s), 5.04(2H, s), 4.12(2H, t, J=6.8 Hz), 4.04(3H, s), 1.67(2H, m),
0.94(3H, t, J=7.3 Hz).
Reference Example 78
6-Amino-9-benzyl-2-(2-methoxyethyl)aminopurine
##STR176##
6-Amino-9-benzyl-2-chloropurine (10 mg, 0.385 mmol) and
2-methoxyethylamine in 2 ml of butanol were heated at 120.degree.
C. for 9 hours in autoclave. The reaction mixture was concentrated
in vacuo to dryness and to the residue was added water. The mixture
was extracted with chloroform and the organic layer was dried on
sodium sulfate and concentrated in vacuo to dryness. The residue
was purified with silica gel chromatography (3%
methanol/chloroform) to give the subject compound (83 mg, yield
72%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.81(1H, s), 7.35-7.26(5H, m),
6.72(2H, br s), 6.18(1H, t, J=4.8 Hz), 5.19(2H, s), 3.45-3.36(4H,
m), 3.24(3H, s).
Reference Example 79
6-Amino-9-benzyl-8-bromo-2-(2-methoxyethylamino)purine
##STR177##
6-Amino-9-benzyl-2-(2-methoxyethyl)aminopurine (70 mg, 0.24 mmol)
and bromine (0.5 ml) were dissolved in methylene chloride (50 ml).
The solution was stirred at room temperature for 1 hour. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated, dried on sodium sulfate and concentrated in
vacuo to dryness. The residue was purified with silica gel
chromatography (1% methanol/chloroform) to give the subject
compound (71 mg, yield 80%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.36-7.23(5H, m), 6.94(2H, br
s), 6.38(1H, t, J=4.8 Hz), 5.18(2H, s), 3.45-3.36(4H, m), 3.23(3H,
s).
Reference Example 80
6-Amino-9-benzyl-8-methoxy-2-(2-methoxyethyl]aminopurine
##STR178##
6-Amino-9-benzyl-8-bromo-2-(2-methoxyethyl)aminopurine (68 mg, 0.18
mmol) was dissolved in 28% sodium methoxide in methanol (30 ml) and
the solution was refluxed on heating under stirring for 4 hours.
The reaction mixture was concentrated in vacuo to dryness and to
the residue was added water. The mixture was extracted with
chloroform and the organic layer was dried on sodium sulfate and
concentrated in vacuo to dryness. The residue was purified with
silica gel chromatography (2% methanol/chloroform) to give the
subject compound (26 mg, yield 44%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.35-7.21(5H, m), 6.36(2H, br
s), 6.01 (1H, t, J=4.8 Hz), 4.98(2H, s), 3.99(3H, s), 3.45-3.36(4H,
m), 3.23 (3H, S).
Reference Example 81
6-Amino-9-benzyl-8-methoxy-2-(2-ethoxyethoxy)purine
##STR179##
6-Amino-9-benzyl-2-chloropurine (500 mg, 1.93 mmol) was dissolved
in 40 ml of sodium 2-ethoxyethoxy in 2-ethoxyethanol and the
solution was heated at 100.degree. C. for 6 hours. The reaction
mixture was concentrated in vacuo to dryness and to the residue was
added water. The mixture was extracted with chloroform and the
organic layer was washed with water, dried on sodium sulfate and
concentrated in vacuo to dryness. The residue was purified with
silica gel chromatography (2% methanol/chloroform) to give the
subject compound (410 mg, yield 68%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 8.05(1H, s), 7.38-7.26(5H, m),
7.24(2H, br s), 5.26(2H, s), 4.32(2H, t, J=4.8 Hz), 3.65(2H, t,
J=4.8 Hz), 3.47(2H, q, J=7.0 Hz), 1.11(3H, t, J=7.0 Hz).
Reference Example 82
6-Amino-9-benzyl-8-bromo-2-(2-ethoxyethoxy)purine
##STR180##
6-Amino-9-benzyl-2-(2-ethoxyethoxy)purine (300 mg, 0.96 mmol) and
bromine (2.0 ml) were dissolved in methylene chloride (50 ml). The
solution was stirred at room temperature for 1 hour. Aqueous sodium
thiosulfate was added to the reaction mixture. The organic layer
was separated, dried on sodium sulfate and concentrated in vacuo to
dryness. The residue was purified with silica gel chromatography
(1% methanol/chloroform) to give the subject compound (256 mg,
yield 68%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.47(2H, br s), 7.39-7.23(5H,
m), 5.26(2H, s), 4.32(2H, t, J=4.8 Hz), 3.65(2H,t, J=4.8 Hz),
3.47(2H, q, J=7.0 Hz), 1.11(3H, t, J=7.0 Hz).
Reference Example 83
6-Amino-9-benzyl-2-(2-ethoxyethoxyl-8-methoxypurine
##STR181##
6-Amino-9-benzyl-8-bromo-2-(2-ethoxyethoxy)purine (206 mg, 0.18
mmol) was dissolved in 1N sodium hydroxide in methanol (20 ml) and
the mixture was refluxed on heating under stirring for 2 hours. The
reaction mixture was concentrated in vacuo to dryness, and to the
residue was added water. The solution was extracted with chloroform
and the organic layer was dried on sodium sulfate and concentrated
in vacuo to dryness. The residue was purified with silica gel
chromatography (2% methanol/chloroform) to give the subject
compound (123 mg, yield 68%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.36-7.23(5H, m), 6.89(2H, br
s), 5.04(2H, s), 4.29(2H, t, J=4.6 Hz), 4.05(3H, s), 3.64(2H, t,
J=4.6 Hz), 3.47(2H, q, J=7.0 Hz), 1.11(3H, t, J=7.0 Hz).
Reference Example 84
6-Amino-2-chloro-9-(4-fluorobenzyl)purine
##STR182##
6-Amino-2-chloropurine (5.02 g) and potassium carbonate (5 g, 36
mmol) were suspended in DMF (200 ml) and thereto was added
4-fluorobenzyl chloride (5 ml, 42 mmol). The mixture was stirred at
room temperature for 3 hours. The reaction mixture was concentrated
in vacuo to dryness, and to the residue was added water. The
mixture was extracted with chloroform and the organic layer was
washed with water, dried on sodium sulfate and concentrated in
vacuo to dryness. The residue was purified with silica gel
chromatography (2% methanol/chloroform) to give the subject
compound (162 g).
.sup.1 H-NMR(DMSO-dE) .delta.: 8.25(1H, s), 7.80(2H, br s),
7.37(5H, m), 7.18(2H, m), 5.33(2H, s).
Reference Example 85
6-Amino-9-(4-fluorobenzyl)-2-(2-methoxyethoxy)purine
##STR183##
6-Amino-9-(4-fluorobenzyl)purine (100 mg, 0.36 mmol) was dissolved
in 30 ml of sodium 2-mehoxyethoxide in 2-methoxyethanol and the
solution was heated at 20.degree. C. for 3 hours. The reaction
mixture was concentrated in vacuo to dryness and to the residue was
added water. The mixture was extracted with chloroform and the
organic layer was washed with water, dried on sodium sulfate and
concentrated in vacuo to dryness. The residue was purified with
silica gel chromatography (2% methanol/chloroform) to give the
subject compound (109 mg, yield 95%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 8.05(1H, s), 7.40 (2H, m),
7.24(2H, br s), 7.17(2H, m), 5.25(2H, s), 4.33(2H, t, J=4.4 Hz),
3.62(2H, t, J=4.4 Hz), 3.29(3H, s).
Reference Example 86
6-Amino-8-bromo-9-(fluorobenzyl)-2-(2-methoxyethoxy)purine
##STR184##
6-Amino-9-(4-fluorobenzyl)-2-(2-methoxyethoxy)purine (mg, 0.96
mmol) and bromine (1.0 ml) were dissolved in methylene chloride (20
ml). The solution was stirred at room temperature for 1 hour.
Aqueous sodium thiosulfate was added to the reaction mixture. The
organic layer was separated, dried on sodium sulfate and
concentrated in vacuo to dryness. The residue was purified with
silica gel chromatography (1% methanol/chloroform) to give the
subject compound (79 mg, yield 69%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.46(2H, br s), 7.31(2H, m),
7.19(2H, m), 5.24(2H, s), 4.34(2H, t, J=4.6 Hz), 3.62(2H,t, J=4.6
Hz), 3.29(3H, s).
Reference Example 87
6-Amino-9-(4-fuluorobenzyl)-8-methoxy-2-(2-methoxyethoxy)purine
##STR185##
6-Amino-8-bromo-9-(4-fluorobenzyl)-2-(2-methoxyethoxy)purine (70
mg, 0.18 mmol) was dissolved in 1N sodium hydroxide in methanol (20
ml) and the solution was refluxed on heating under stirring for 2
hours. The reaction mixture was concentrated in vacuo to dryness,
and to the residue was added water. The mixture was extracted with
chloroform and the organic layer was dried on sodium sulfate and
concentrated in vacuo to dryness. The residue was purified with
silica gel chromatography (2% methanol/chloroform) to give the
subject compound (57 mg, yield 93%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.31(2H, m), 7.16(2H, m),
6.87(2H, br s), 5.02(2H, s), 4.30(2H, t, J=4.6 Hz), 4.05(3H, s),
3.61(2H, t, J=4.6 Hz), 3.29(3H, s).
Reference Example 88
5-Amino-4-cyano-1-(4-fluorobenzyl)-2-hydroxyimidazole
##STR186##
4-Fluorobenzylisocyanate (1.37 g, 10 mmol) and
N,N-diisopropylethylamine (1.29 g, 10 mmol) were added to
aminomalononitrile p-toluenesulfonate (2.53 g, 10 mmol) suspended
in tetrahydrofuran (50 ml). The mixture was stirred at room
temperature for 24 hours and then the solvent was removed in vacuo.
To the residue was added ethyl acetate and the solution was washed
and the organic layer was extracted with 1N aqueous sodium
hydroxide. The extract was neutralized with 10% aqueous potassium
hydrogen sulfide and extracted with ethyl acetate. The organic
layer was dried on sodium sulfate and the solvent was removed in
vacuo. The residue was purified with silica gel chromatography (1%
methanol/chloroform) to give the subject compound (1.93 g, yield
89%).
.sup.1 H-NMR (DMSO-d.sub.6): 9.93(1H, br s), 7.31(2H, m), 7.15(2H,
m), 6.53(2H, br s), 4.75(2H, s).
Reference Example 89
6-Amino-9-(4-fluorobenzyl)-8-hydroxy-2-thiopurine
##STR187##
5-amino-4-cyano-1-(4-fluorobenzyl)-2-hydroxyimidazole (1.90 g, 8.79
mmol) was suspended in tetrahydrofuran (50 ml) and to the
suspension was dropped benzoylisothiocyanate (2.87 g, 17.6 mmol).
After stirring at room temperature for 8 hours, the solvent was
removed in vacuo and to the residue was added ether. The crystals
were filtered and refluxed in a mixture of tetrahydrofuran and 1N
aqueous sodium hydroxide for 40 hours and neutralized with 10%
aqueous potassium hydrogen sulfide. The crystals were harvested by
filtration to give the subject compound (1.22 g, yield: 48%).
.sup.1 H NMR (.delta., DMSO-d.sub.6): 10.14(1H, s), 7.31(2H, m),
7.15(2H, m), 6.83(2H, s), 4.84(2H, s).
Reference Example 90
6-Amino-9-benzyl-2,8-dihydroxypurine
##STR188##
6-Amino-9-benzyl-8-bromo-2-methoxypurine (75 mg, 0.224 mmol) in
concentrated hydrochloric acid (15 ml) was refluxed under heating
for 5 hours. The reaction mixture was made basic by 28% aqueous
ammonia, the crystals were filtered, washed with water and purified
by silica gel chromatography (0.2%aqueous ammonia-5%
methanol/chroloform) to give the subject compound (12 mg, yield
21%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 9.64(2H, br s), 7.34-7.22(6H,
m), 6.51(2H, br s), 4.78(2H, s).
Reference Example 91
2,6-Diamino-9-benzyl-8-purinol
##STR189##
2,6-Diamino-9-benzyl-8-bromopurine (400 mg, 1.25 mmol) in
concentrated hydrochloric acid (20 ml) was refluxed under heating
for 5 hours. The reaction mixture was made basic by 28% aqueous
ammonia, the crystals were filtered, washed with water and dried to
give the subject compound (138 mg, yield 43%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 9.63 (1H, br s), 7.34-7.22 (5H,
m), 6.02 (2H, br s), 5.74 (2H, br s), 4.81 (2H, s).
Reference Example 92
6-Amino-9-benzyl-2-(2-aminoethyl)thiopurine
##STR190##
To sodium hydride (300 mg, 7.5 mmol, 60% in mineral oil) were added
DMF (10 ml), 2-aminoethanethiol (620 mg, 8 mmol) and
6-amino-9-benzyl-2-chloropurine (200 mg, 0.77 mmol) in order. The
mixture was stirred at 100.degree. C. for 3 hours. After addition
of saturated brine the reaction mixture was extracted with
chloroform and the organic layer was dried on magnesium sulfate and
concentrated in vacuo to dryness. The residue was purified with
silica gel chromatography (10% methanol/chloroform) to give the
subject compound (126 mg, yield 54%).
.sup.1 H NMR (CDCl.sub.3) .delta.: 7.64 (s, 1H), 7.31-7.26 (m, 5H),
5.53 (br s, 2H), 5.29 (s, 2H), 3.26 (t, 2H, J=6.0 Hz), 3.02 (t, 2H,
J=6.3 Hz).
Reference Example 93
6-Amino-9-benzyl-2-(2-dimethylaminoethyl)thiopurine
##STR191##
To sodium hydride (600 mg, 15 mmol 60% in mineral oil) were added
DMF (10 ml), 2-dimethylaminoethanethiol (1.3 g, 9.2 mmol) and
6-amino-9-benzyl-2-chloropurine (100 mg, 0.39 mmol) in order. The
mixture was stirred at 100.degree. C. for 10 hours. After addition
of saturated brine the reaction mixture was extracted with
chloroform and the organic layer was dried on magnesium sulfate and
concentrated in vacuo to dryness. The residue was purified with
silica gel chromatography (10% methanol/chloroform) to give the
subject compound (24 mg, yield 21%).
.sup.1 H NMR (CDCl.sub.3) .delta.: 7.63 (s, 1H), 7.36-7.26 (m, 5H),
5.55 (br s, 2H), 5.30 (s, 2H), 3.29 (t, 2H, J=7.6 Hz), 2.68 (t, 2H,
J=7.9 Hz), 2.30 (s, 6H), 1.80 (br s, 2H).
Reference Example 94
3-(6-Amino-9-benzyl-2-purinyl)thiopropionic Acid
##STR192##
To sodium hydride (300 mg, 7.5 mmol 60% in mineral oil) were added
DMF (10 ml), 3-mercaptopropionic acid (1 ml, 11 mmol) and
6-amino-9-benzyl-2-chloropurine (200 mg, 0.77 mmol) in order. The
mixture was stirred at 100.degree. C. for 5 hours. After addition
of saturated brine, the reaction mixture was acidified with 2N
hydrochloric acid and extracted with chloroform. The organic layer
was dried on magnesium sulfate and concentrated in vacuo to
dryness. The residue was purified with silica gel chromatography
(4% methanol/chloroform) to give the subject compound (120 mg,
yield 47%).
.sup.1 H NMR (DMSO-d.sub.6) .delta.: 12.26 (br s, 1H); 8.16 (s,
1H), 7.39-7.17 (m, 5H), 5.29 (s, 2H), 3.22 (t, 2H, J=7.2 Hz), 2.66
(t, 2H, J=6.9 Hz).
Reference Example 95
2-(2-Acetylaminoethyl)thio-6-amino-9-benzylpurine
##STR193##
To 6-amino-9-benzyl-2-(2-aminoethyl)thiopurine (50 mg, 0.17 mmol)
suspended in 2 ml of dichloromethane was added triethylamine (30
ml, 0.2 mmol) and then acetic acid anhydride (20 ml, 0.2 mmol)
under ice cooling. After 1 hour the mixture warmed to room
temperature. Thereto was added saturated brine. The mixture was
extracted with chloroform and the organic layer was dried on
magnesium sulfate and concentrated in vacuo to dryness. The residue
was purified with silica gel chromatography (1%
methanol/chloroform) to give the subject compound (34 mg, yield
66%).
.sup.1 H NMR (CDCl.sub.3) .delta.: 7.67 (s, 1H), 7.34-7.26 (m, 5H),
6.17 (br s, 3H), 5.31 (s, 2H), 3.59 (q, 2H, J=5.6 Hz), 3.31 (t, 2H,
J=5.2 Hz).
Reference Example 96
Methyl-3-(6-amino-9-benzyl-2-purinyl)thiopropionate
##STR194##
To 3-(6-amino-9-benzyl-2-purinyl) thiopropionic acid (100 mg, 0.30
mmol) suspended in 10 ml of chloroform was added thionylchloride
(0.14 ml, 2 mmol). After refluxing under heating for 1 hour
methanol was dropped to the mixture under ice cooling. The solvent
was removed in vacuo and the residue was purified by silica gel
chromatography (1% methanol/chloroform) to give the subject
compound (70 mg, yield 68%).
.sup.1 H NMR (CDCl.sub.3) .delta.: 7.65 (s, 1H), 7.36-7.26 (m, 5H),
5.67 (br s, 2H), 5.28 (s, 2H), 3.69 (s, 3H), 3.39 (t, 2H, J=7.3
Hz), 2.82 (t, 2H, J=7.6 Hz).
Reference Example 97
N-Methyl-(6-amino-9-benzyl-2-purinyl)thioacetamide
##STR195##
Sodium hydride (320 mg, 8 mmol 60% in mineral oil) was washed with
hexane. Thereto were added DMF (10 ml),
2-mercapto-N-methylacetamide (1 ml) and
6-amino-9-benzyl-2-chloropurine (200 mg, 0.77 mmol) in order. The
mixture was stirred at 100.degree. C. for 8 hours. After addition
of saturated brine the reaction mixture was extracted with ethyl
acetate. The organic layer was dried on magnesium sulfate and the
solvent was removed in vacuo. The residue was purified with silica
gel chromatography (3% methanol/chloroform) to give the subject
compound (158 mg, yield 60%).
.sup.1 H NMR (DMSO-d.sub.6) .delta.: 8.14 (s, 1H), 7.87 (br s, 1H),
7.33 (m, 5H), 5.30 (s, 2H), 3.75 (s, 2H), 2.53 (d, 3H, J=4.6
Hz).
Reference Example 98
3-(6-Amino-9-benzyl-2-purinyl)thio-1-propanol
##STR196##
Sodium hydride (600 mg, 15 mmol 60% in mineral oil) were added DMF
(10 ml), 3-mercapto-1-propanol (1 ml, 12 mmol) and
6-amino-9-benzyl-2-chloropurine (200 mg, 0.77 mmol) in order. The
mixture was stirred at 100.degree. C. for 2 hours. After addition
of saturated brine the reaction mixture was extracted with ethyl
acetate. The organic layer was dried on magnesium sulfate and the
solvent was removed in vacuo. The residue was purified with silica
gel chromatography (1% methanol/chloroform) to give the subject
compound (30 mg, yield 12%).
.sup.1 H NMR (CDCl.sub.3) .delta.: 7.62 (s, 1H), 7.37-7.26 (m, 5H),
5.59 (br s, 2H), 5.29 (s, 2H), 3.76 (m, 3H), 3.33 (t, 2H, J=6.3
Hz), 1.96 (m, 2H).
Reference Example 99
3-(6-Amino-9-benzyl-2-purinylithio-1-propanethiol
##STR197##
Sodium hydride (600 mg, 15 mmol 60% in mineral oil) were added DMF
(10 ml), 1,3-propanedithiol (1 ml, 10 mmol) and
6-amino-9-benzyl-2-chloropurine (200 mg, 0.77 mmol) in order. The
mixture was stirred at 100.degree. C. for 2 hours. After addition
of saturated brine the reaction mixture was extracted with ethyl
acetate. The organic layer was dried on magnesium sulfate and the
solvent was removed in vacuo. The residue was purified with silica
gel chromatography (1% methanol/chloroform) to give the subject
compound (135 mg, yield 53%).
.sup.1 H NMR (CDCl.sub.3) .delta.: 7.65 (s, 1H), 7.36-7.26 (m, 5H),
5.56 (br s, 2H), 5.31 (s, 2H), 3.28 (t, 2H, J=6.6 Hz), 2.65 (m,
2H), 2.05 (m, 2H), 1.40 (t, 1H, J=7.9 Hz).
Reference Example 100
6-Amino-9-benzyl-2-(2-phenylethyl)thiopurine
##STR198##
Sodium hydride (300 mg, 7.5 mmol 60% in mineral oil) were added DMF
(10 ml), 2-phenylethanethiol (1 ml, 7 mmol) and
6-amino-9-benzyl-2-chloropurine (100 mg, 0.39 mmol) in order. The
mixture was stirred at 100.degree. C. for 10 hours. After addition
of saturated brine the mixture was extracted with ethyl acetate.
The organic layer was dried on magnesium sulfate and the solvent
was removed in vacuo. The residue was purified with silica gel
chromatography (1% methanol/chloroform) to give the subject
compound (51 mg, yield 37%).
.sup.1 H NMR (CDCl.sub.3) .delta.: 7.65 (s, 1H), 7.32-7.24 (m, 5H),
5.57 (br s, 2H), 5.33 (s, 2H), 3.39 (dd, 2H, J=10.6, 7.6 Hz), 3.07
(dd, 2H, J=15.8, 7.2 Hz).
Reference Example 101
6-Amino-9-benzyl-2-(2-hydroxyethoxy)purine
##STR199##
To sodium (74 mg, 3.2 mmol) in 5 ml of ethylene glycol were added
6-amino-9-benzyl-2-chloropurine (157 mg, 0.58 mmol). The mixture
was heated at 100.degree. C. for 4 hours and concentrated in vacuo
to dryness. To the residue was added water and the mixture was
extracted with chloroform. The organic layer was dried on magnesium
sulfate and the solvent was removed in vacuo. The residue was
purified with silica gel chromatography (3% methanol/chloroform) to
give the subject compound (121 mg, yield 70%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 8.04(1H, s), 7.32(5H, m),
7.22(2H, br s), 5.25(2H, s), 4.81(1H, t, J=5.3 Hz), 4.22(2H, t,
J=4.9 Hz), 3.68(2H, q, J=5.3 Hz).
Reference Example 102
6-Amino-9-benzyl-8-bromo-2-(2-hydroxyethoxy)purine
##STR200##
6-Amino-9-benzyl-2-(2-methoxyethoxy)purine (100 mg, 0.36 mmol) and
bromine (0.25 ml) were dissolved in methylene chloride (100 ml).
The solution was stirred at room temperature for 6 hours. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated, dried on sodium sulfate and the solvent was
removed in vacuo. The residue was purified with silica gel
chromatography (2% methanol/chloroform) to give the subject
compound (55 mg, yield 43%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.44(2H, br s), 7.36-7.23(5H,
m), 5.25(2H, s), 4.82(1H, t, J=5.6 Hz), 4.22(2H, t, J=5.0 Hz),
3.66(2H, q, J=5.0 Hz).
Reference Example 103
6-Amino-9-benzyl-2-(2-hydroxyethoxyl)-8-methoxypurine
##STR201##
6-Amino-9-benzyl-8-bromo-2-(2-methoxyethoxyl)purine (130 mg, 0.36
mmol) in methanol (50 ml) was dissolved in 28% sodium
methoxide/methanol (3 ml) and the solution was refluxed on heating
under stirring for 10 hours. The reaction mixture was concentrated
in vacuo to dryness, and to the residue was added saturated brine.
The mixture was extracted with chloroform and the organic layer was
dried on sodium sulfate, followed by removal of the solvent. The
residue was purified with silica gel chromatography (3%
methanol/chloroform) to give the subject compound (78 mg, yield
69%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.26(5H, m), 6.86(2H, br s),
5.03(2H, s), 4.78(1H, t, J=5.6 Hz), 4.18(2H, t, J=5.0 Hz), 4.04(3H,
s), 3.66(2H, m).
Reference Example 104
6-Amino-9-benzyl-2-(3-hydroxypropoxy)purine
##STR202##
Sodium (80 mg, 3.5 mmol) in 3 ml of 1,3-propanediol was added
6-amino-9-benzyl-2-chloropurine (235 mg, 0.90 mmol). The mixture
was heated at 100.degree. C. for 3 hours. The reaction mixture was
concentrated in vacuo to dryness. To the residue was added water
and the mixture was extracted with chloroform. The organic layer
was dried on sodium sulfate and the solvent was removed. The
residue was purified with silica gel chromatography (3%
methanol/chloroform) to give the subject compound (137 mg, yield
51%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 8.03(1H, s), 7.32(5H, m),
7.21(2H, br s), 5.25(2H, s), 4.51(1H, t, J=4.9 Hz), 4.26(2H, t,
J=6.3 Hz), 3.52(2H, q, J=5.6 Hz), 1.81(2H, m).
Reference Example 105
6-Amino-9-benzyl-8-bromo-2-(3-hydroxypropoxy)purine
##STR203##
6-Amino-9-benzyl-2-(3-hydroxypropoxy)purine (210 mg, 0.7 mmol) and
bromine (0.5 ml) were dissolved in methylene chloride (200 ml). The
solution was stirred at room temperature for 4 hours. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated, dried on magnesium sulfate and concentrated in
vacuo to dryness. The residue was purified with silica gel
chromatography (5% methanol/chloroform) to give the subject
compound (143 mg, yield 54%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.45(2H, br s), 7.32(5H, m),
5.25(2H, s), 4.52(1H, t, J=5.0 Hz), 4.26(2H, t, J=6.6 Hz), 3.52(2H,
q, J=5.6 Hz), 1.81(2H, m).
Reference Example 106
6-Amino-9-benzyl-2-(3-hydroxypropoxy)-8-methoxypurine
##STR204##
6-Amino-9-benzyl-8-bromo-2-(3-hydroxypropoxy)purine (140 mg, 0.37
mmol) in methanol (50 ml) was dissolved in 28%. sodium
methoxide/methanol (3 ml) and the solution was refluxed on heating
under stirring for 10 hours. The reaction mixture was concentrated
in vacuo to dryness and to the residue was added saturated brine.
The mixture was extracted with chloroform and the organic layer was
dried on sodium sulfate and the solvent was removed. The residue
was purified with silica gel chromatography (3%
methanol/chloroform) to give the subject compound (88 mg, yield
72%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.31(5H, m), 6.86(2H, br s),
5.04(2H, s), 4.50(1H, t, J=5.0 Hz), 4.22(2H, t, J=6.6 Hz), 4.03(3H,
s), 3.52(2H, m), 1.80(2H, m).
Reference Example 107
6-Amino-9-benzyl-2-(3-ethoxypropoxy)purine
##STR205##
To sodium (150 mg, 6.5 mmol) in 5 ml of 3-ethoxypropanol were added
6-amino-9-benzyl-2-chloropurine (500 mg, 1.93 mmol) and DMF (10
ml). The mixture was heated at 120.degree. C. for 1 hour. The
reaction mixture was concentrated in vacuo to dryness. To the
residue was added water and the mixture was extracted with
chloroform. The organic layer was dried on sodium sulfate and the
solvent was removed. The residue was purified with silica gel
chromatography (2% methanol/chloroform) to give the subject
compound (481 mg, yield 76%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 8.04(1H, s), 7.34-7.25(5H, m),
7.24(2H, br s), 5.27(2H, s), 4.27(2H, t, J=6.4 Hz), 3.48(2H, t,
J=6.4 Hz), 3.41(2H, q, J=7.0 Hz), 1.91(2H, m), 1.10(3H, t, J=7.0
Hz).
Reference Example 108
6-Amino-9-benzyl-8-bromo-2-(3-ethoxypropoxy)purine
##STR206##
6-Amino-9-benzyl-2-(3-ethoxypropoxy)purine (354 mg, 1.08 mmol) and
bromine (1.0 ml) were dissolved in methylene chloride (50 ml). The
solution was stirred at room temperature for 2 hours. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated, dried on sodium sulfate and the solvent was
removed. The residue was purified with silica gel chromatography
(1% methanol/chloroform) to give the subject compound (289 mg,
yield 66%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.37(2H, br s), 7.36-7.23(5H,
m), 5.26(2H, s), 4.26(2H, t, J=6.4 Hz), 3.47(2H, t, J=6.4 Hz),
3.40(2H, q, J=7.0 Hz), 1.90(2H, m), 1.09(3H, t, J=7.0 Hz).
Reference Example 109
6-Amino-9-benzyl-2-(3-ethoxypropoxy)-8-methoxypurine
##STR207##
To 6-amino-9-benzyl-8-bromo-2-(3-ethoxypropoxy)purine (250 mg, 0.36
mmol) in 20 ml of methanol was added 1N aqueous sodium hydroxide
(80 ml). The mixture was refluxed on heating under stirring for 2
hours. The reaction mixture was concentrated in vacuo to dryness.
To the residue was added water and the mixture was extracted with
chloroform. The organic layer was dried on sodium sulfate and the
solvent was removed. The residue was purified with silica gel
chromatography (2% methanol/chloroform) to give the subject
compound (187 mg, yield 85%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.36-7.23(5H, m), 6.88(2H, br
s), 5.05(2H, s), 4.23(2H, t, J=6.4 Hz), 4.05(3H, s), 3.47(2H, t,
J=6.4 Hz), 3.38(2H, q, J=7.0 Hz), 1.89(2H, m), 1.10(3H, t, J=7.0
Hz).
Reference Example 110
6-Amino-9-benzyl-2-(4-hydroxybutoxy)purine
##STR208##
Sodium (150 mg, 6.5 mmol) in 5 ml of 1,5-butanediol were added
6-amino-9-benzyl-2-chloropurine (500 mg, 1.93 mmol) and DMF (10
ml). The mixture was heated at 120.degree. C. for 1 hour. The
reaction mixture was concentrated in vacuo to dryness. To the
residue was added water and the mixture was extracted with
chloroform. The organic layer was dried on sodium sulfate and the
solvent was removed. The residue was purified with silica gel
chromatography (3% methanol/chloroform) to give the subject
compound (336 mg, yield 56%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 8.04(1H, s), 7.37-7.25(5H, m),
7.22(2H, br s), 5.26(2H, s), 4.46(1H, t, J=5.3 Hz), 4.22(2H, t,
J=6.6 Hz), 3.43(2H, m), 1.71(2H, m), 1.53(2H, m).
Reference Example 111
6-Amino-9-benzyl-8-bromo-2-(4-hydroxybutoxy)purine
##STR209##
6-Amino-9-benzyl-2-(4-hydroxybutoxy)purine (200 mg, 0.638 mmol) and
bromine (1.0 ml) were dissolved in methylene chloride (50 ml). The
solution was stirred at room temperature for 2 hours. Aqueous
sodium thiosulfate was added to the reaction mixture. The organic
layer was separated, dried on sodium sulfate and the solvent was
removed. The residue was purified with silica gel chromatography
(2% methanol/chloroform) to give the subject compound (213 mg,
yield 85%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.44(2H, br s), 7.39-7.24(5H,
m), 5.26(2H, s), 4.45(1H, t, J=5.1 Hz).sub.1 4.23(2H, t, J=6.6 Hz),
3.44(2H, m), 1.71(2H, m), 1.54(2H, m).
Reference Example 112
6-Amino-9-benzyl-2-(4-hydroxybutoxy)-8-methoxypurine
##STR210##
To 6-amino-9-benzyl-8-bromo-2-(4-hydroxybutoxy)purine (185 mg,
0.472 mmol) in 10 ml of methanol was added 1N aqueous sodium
hydroxide (40 ml). The mixture was refluxed on heating under
stirring for 2 hours. The reaction mixture was concentrated in
vacuo to dryness. To the residue was added water and the mixture
was extracted with chloroform. The organic layer was dried on
sodium sulfate and the solvent was removed. The residue was
purified with silica gel chromatography (2% methanol/chloroform) to
give the subject compound (123 mg, yield 68%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.37-7.23(5H, m), 6.87(2H, br
s), 5.04(2H, s), 4.43(1H, br s), 4.18(2H, t, J=6.4 Hz), 3.43(2H, t,
J=6.6 Hz), 1.68(2H, m), 1.53(2H, m).
Reference Example 113
6-Amino-9-benzyl-2-(2-methoxyethoxy)purine
##STR211##
Sodium (66 mg, 2.9 mmol) in 50 ml of 2-methoxyethanol was added
6-amino-9-benzyl-2-chloropurine (150 mg, 0.58 mmol) and DMF (10
ml). The mixture was heated at 130.degree. C. for 6 hour. The
reaction mixture was concentrated in vacuo to dryness. To the
residue was added water and the mixture was extracted with
chloroform. The organic layer was dried on sodium sulfate and
concentrated in vacuo to dryness. The residue was purified with
silica gel chromatography (3% methanol/chloroform) to give the
subject compound (123 mg, yield 71%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 8.05(1H, s), 7.36-7.27(5H, m),
7.23(2H, br s), 5.26(2H, s), 4.32(2H, t, J=4.6 Hz), 3.61(2H, t,
J=4.6 Hz), 3.28(3H, s).
Reference Example 114
6-Amino-9-benzyl-8-bromo-2-(2-methoxyethoxy)purine
##STR212##
6-Amino-9-benzyl-2-(2-methoxyethoxy)purine (93 mg, 0.31 mmol) and
bromine (1 ml) were dissolved in methylene chloride (100 ml). The
solution was stirred at room temperature for 1 hour. Aqueous sodium
thiosulfate was added to the reaction mixture. The organic layer
was separated, dried on sodium sulfate and concentrated in vacuo to
dryness. The residue was purified with silica gel chromatography
(1% methanol/chloroform) to give the subject compound (75 mg, yield
64%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.46(2H, br s), 7.38-7.23(5H,
m), 5.25(2H, s), 4.33(2H, t, J=4.6 Hz), 3.61(2H, t, J=4.6 Hz),
3.28(3H, s).
Reference Example 115
6-Amino-9-benzyl-8-methoxy-2-(2-methoxyethoxy)purine
##STR213##
6-Amino-9-benzyl-8-bromo-2-(2-methoxyethoxy)purine (69 mg, 0.18
mmol) in methanol (50 ml) was dissolved in 28% sodium
methoxide/methanol (1 ml) and the mixture was refluxed on heating
under stirring for 5 hours. The reaction mixture was concentrated
in vacuo to dryness, and to the residue was added water. The
solution was extracted with chloroform and the organic layer was
dried on sodium sulfate and concentrated in vacuo to dryness. The
residue was purified with silica gel chromatography (3%
methanol/chloroform) to give the subject compound (26 mg, yield
43%).
.sup.1 H-NMR(DMSO-d.sub.6) .delta.: 7.36-7.23(5H, m), 6.88(2H, br
s), 5.04(2H, s), 4.29(2H, t, J=4.6 Hz), 4.04(3H, s), 3.60(2H, t,
J=4.6 Hz), 3.28(3H, s).
INDUSTRIAL APPLICABILITY
According to the present invention an interferon inducer containing
a compound of the present invention as an active agent is provided.
The interferon inducer of the present invention has inducing and
activating activity for biosynthesis of interferon and therefore,
is useful as therapeutic agents based on biological activities of
interferon, such as antiviral activity, preventing activity of cell
growth, immune modulation etc., that is, therapeutic agents for
virus infected diseases (e.g. hepatitis C, hepatitis B), anticancer
agents and agents for immunologic diseases.
* * * * *